Immunohistochemical detection of Fas antigen in oral epithelia.

A monospecific and high titer polyclonal antibody, designated as Fas D, raised against synthetic polypeptides selected from a part of the human Fas antigen (aa 104-114), was used to identify the Fas antigen in human oral epithelia in normal and pathological states. The human gingival proteins had been extracted and analysed by an ABC (avidin-biotin complex) immunoblotting technique. The antibody interacted with a single band in gingival proteins with an estimated molecular weight of 35,000, which is in good agreement with that calculated from amino acid sequences of the human Fas antigen. Using an indirect immunohistochemical method, the antibody localized on the stratum spinosum and the basal part of the stratum corneum of normal human gingiva. Specimens obtained from patients with odontogenic keratocysts, leukoplakia, lichen planus, and squamous cell carcinoma were also stained with the antibody. The pattern of the Fas antigen distribution in oral stratified squamous epithelia was, with some overlapping, characteristic for each disease.


The protein phosphatase inhibitors okadaic acid and calyculin A induce apoptosis in human osteoblastic cells.

To determine whether protein phosphorylation and dephosphorylation can affect apoptosis in osteoblastic cells, we examined the effects of okadaic acid (OA) and calyculin A (CA) on cultured human osteoblastic cells Saos-2 and MG63, and mouse osteoblastic MC3T3-E1 cells. After reaching confluence, these cells were exposed to varying concentrations of OA or CA. OA and CA induced cell death in all three cell lines in a dose- and time-dependent manner. Marked nuclear condensation and fragmentation of chromatin were also observed in these cells by using the Hoechst 33342 stain. DNA ladder formation, a hallmark of apoptosis, was detected in Saos-2 and MG63 cells, but not in MC3T3-E1 cells by treatment of OA or CA. In the Saos-2 cells, OA- and CA-induced DNA ladder formation was dose-dependent with maximal effect at concentrations of 10 and 2 nM, respectively, and was time-dependent from 14 to 48 h. DNA ladder formation in response to OA and CA was revealed by using conventional ethidium bromide staining of electrophoresed DNA without using autoradiography. Beyond the maximal effects at the respective concentrations, however, cell death did not indicate DNA laddering, suggesting that phosphatase activity may be required for ladder formation. Our results indicate that apoptosis in the cultured osteoblastic cells is induced by moderate inhibition of PP-1 or PP-2A based on the known selectivity of okadaic acid and of calyculin A.


Immunohistochemistry of Fas antigen --- Detection of Fas antigen in oral leukoplakia, lichen planus, and squamous cell carcinoma

抗 Fas 抗体 (Fas D) を用いた免疫組織化学的研究 --- 口腔白板症，扁平苔せん，扁平上皮癌における Fas 抗原の検出

Japanese Journal of Oral & Maxillofacial Surgery


Detection of Fas antigen in oral squamous cell carcinoma

口腔扁平上皮癌における Fas 抗原の検出


Immunohistochemical and immunoblotting identification of protein phosphatase 1g1 in rat salivary glands.


Transblot identification of biotin-containing proteins in rat salivary glands.

Biochemistry and Molecular Biology International


Identification of Fas antigen in human gingiva.

The Fas antigen is a cell-surface glycoprotein that mediates apoptosis from the cell surface into the cytoplasm. Polyclonal antibody (Fas D) was raised against a synthetic polypeptide selected from the extracellular part of the human Fas antigen (amino acid residues 104-114) and was used to detect the Fas antigen in human gingiva. Biopsy specimens of human gingiva were prepared, and the paraffin sections were reacted with the Fas D antibody by an immunohistochemical method. The antibody localized to the prickle-cell layer and to granular layer keratinocytes of human gingiva. Proteins were also prepared from human gingiva and subjected to SDS-PAGE, followed by Western-blotting analysis with the Fas D antibody. The antibody interacted with a band corresponding to an estimated molecular weight of 35 kDa. The incidence of the immunoreactive 35-kDa protein was detected in the gingiva of 90% of the 20 individuals examined. The Fas antigen detected in human gingiva may be related to the physiological turnover of oral mucosa.


Protein phosphatase inhibitors, okadaic acid and calyculin A, induce alkaline phosphatase activity in osteoblastic cells derived from newborn mouse calvaria

To determine whether protein phosphatases can affect bone regulation, we examined the effects of okadaic acid (OA) and calyculin A (CA), specific inhibitors of protein phosphatases type 1 and type 2A, on alkaline phosphatase activity of mouse osteoblastic cells. Clone MC3T3-E1 cells were cultured with varying concentrations of OA and CA. OA and CA stimulated alkaline phosphatase (ALP) activity in the cells in dose-dependent fashion with a maximal effect at concentrations of 5 nM and 2 nM, respectively. The properties of OA-induced and native ALP in the cells were the same and they were liver-bone-kidney type. These results show that protein phosphatase inhibitors stimulate bone formation in vitro and that phosphorylation and dephosphorylation of specific proteins in the cells may be involved in bone regulation in vivo as well.


Identification of biotin-containing proteins in rat salivary glands

Journal of the Japan Salivary Gland Society


Testis-specific Mak protein kinase is expressed specifically in the meiotic phase in spermatogenesis and is associated with a 210-kilodalton cellular phosphoprotein

The mak gene encodes a new protein kinase distantly related to cdc2 kinase, and its transcripts are expressed exclusively in testicular germ cells at and after meiosis (H. Matsushime, A. Jinno, N. Takagi, and M. Shibuya, Mol. Cell. Biol. 10:2261-2268, 1990). In this study, we prepared a series of antibodies against synthetic peptides and fusion products of the mak gene and characterized the subcellular localization, protein kinase activity, and association with other cellular proteins of Mak. Mak products were identified as 66- and 60-kDa proteins that specifically appeared in rat testes after puberty. Testicular germ cell fractionation revealed that Mak products were most abundant in the fraction of the late pachytene stage and that their levels were dramatically decreased in postmeiotic haploid cells. Mak products were localized mostly in the cytoplasm as a soluble form. [35S]methionine labelling demonstrated that Mak products were associated with a 210-kDa cellular protein; in an in vitro kinase assay with immunoprecipitates of Mak products, the 210-kDa cellular protein was efficiently phosphorylated on serine and threonine residues. Furthermore, in a testicular cell culture system with 32Pi, the 210-kDa molecule associated with Mak was phosphorylated in vivo on serine and threonine residues. These results strongly suggest that the Mak complex may play a role in meiosis during spermatogenesis and that a phosphorylated 210-kDa protein is one of the physiological substrates for this protein kinase.


Expression of PP2A B regulatory subunit b isotype in rat testis

We isolated a rat cDNA encoding part of the beta-isotype of the B regulatory subunit (BR beta) of protein phosphatase 2A (PP2A). The isolated cDNA encoded the region corresponding to amino acids positions 8(R) to 177(N) of human BR beta. The identities of the nucleotide and amino acid sequences of the rat and human BR beta s were 95.7% and 100%, respectively. The BR beta mRNA was specifically expressed in rat brain and testis, the lengths of mRNAs in these two organs being different. In the testis, the BR beta mRNA was first detected 40 days after birth, increasing gradually thereafter, and was expressed specifically in elongated spermatids, while mRNA of the alpha-isotype (BR alpha) was expressed equally in all spermatogenic cells. After meiosis, round spermatids change morphologically to elongated spermatids. BR beta may regulate the activity of the PP2A catalytic subunit in spermatids, and be involved in spermatogenic maturation, especially spermatid elongation.


Expression of protein phosphatase 1 and 2A in rat testis


Protein phosphatase 1g1 is associated with nuclei of meiotic cells in rat testis

Biochemical and Biophysical Research Communications


Detection of endogeneous avidin-interacting proteins in rat liver cells by transblot and electron microscope


Purification of human sperm antigen interacting with antibodies in serum from an infertile woman


Identification of pyruvate carboxylase in 3T3-L1 cells by transblot and its correlation with cell differentiation

The 3T3-L1 preadipocytes treated with insulin, dexamethasone and 3-methyl-1-isobutylxanthine (IBMX) two days before reaching monolayer undergo differentiation into adipocytes. Cell lysates were prepared from these cells under various conditions and analyzed by SDS-PAGE and transblot. Peroxidase-conjugated avidin used to detect endogenous proteins interacted strongly with a protein with an estimated molecular weight of 120 kDa, corresponding to pyruvate carboxylase, in the differentiated 3T3-L1 cells. On the other hand, this protein was not detected in undifferentiated 3T3-L1 cells.


Evaluation of spermatocidal activity of water soluble gossypol on the sperm of Arbacia punctulata and Spisula solidissima


Identification and expression of boar sperm proteins that interact with antibodies in serum from an infertile woman

Serum designated as IS obtained from a young healthy infertile woman induced a head-to-head agglutination of ejaculated boar sperm. The immunoglobulin G (IgG) prepared from IS localized to the acrosomal region of the sperm head obtained from the corpus and cauda epididymis as determined by an indirect immunofluorescent method. The IgG interacted with a boar sperm protein with an estimated molecular weight of 45-kDa, determined by sodium dodecyl sulfate-polyacrylamide gel electrophoretic (SDS-PAGE) immunoblotting technique. However, the IgG did not interact with proteins extracted from sperm obtained from the testis and caput epididymis or from non-gonadal tissues including liver, kidney, spleen, muscle and serum. The IgG interacted with additional proteins of about 75- and 38-kDa present in the corpus and cauda epididymal fluids but not those in the caput epididymal fluid. The staining intensity of the 75-kDa band was reduced and that of the 38-kDa was nullified with ejaculated seminal plasma proteins. The interacting proteins were adsorbed when chromatographed on Concanavalin A Sepharose column, suggesting that they are glycoproteins.


Differntial effects of Epidermal Growth Factor on the differentiation of type A spermatogonia in adult mouse cryptorchid testes in vitro

The effect of epidermal growth factor (EGF) on testicular germ cell differentiation was investigated. Testicular fragments from surgically prepared cryptorchid testes of adult mice were cultured for 9 days in serum-free media containing various concentrations of EGF. Histological sections of testis were examined under a light microscope and each type of germ cell and mitotic cell in the seminiferous tubules was counted per 1000 Sertoli cells. EGF at concentrations ranging from 100 to 200 ng/ml induced differentiation of type A spermatogonia. The observed maximal stimulatory activity of EGF at a concentration of 100 ng/ml was 30% of the positive control cultures treated with calf serum. EGF at concentrations ranging from 1 to 100 ng/ml significantly inhibited the mitotic activity of FSH, FSH plus retinol, or FSH plus fetuin on type A spermatogonia and their differentiation. The number of type A spermatogonia in testes cultured with FSH, FSH plus retinol, or FSH plus fetuin decreased when EGF was added. On the other hand, EGF stimulated the differentiation of type A spermatogonia induced with fetuin but did not influence retinol-induced differentiation. It is proposed that EGF inhibits testicular germ cell differentiation by blocking the proliferation of type A spermatogonia stimulated by FSH.


Identification of the sperm antigen interacting with antibodies in serum from an infertile woman

Serum (IS) obtained from an infertile woman induced head-to-head agglutination of human sperm. The immunoglobulin G (IgG) fraction of the IS was prepared by ammonium sulfate fractionation and DEAE cellulose chromatography. The IgG localized to the post-acrosomal region of the sperm, determined by indirect immunofluorescence and interacted with a human sperm protein with an estimated Mr of 80 kDa, determined by immunoblotting. The identity of the interacting sperm antigen was verified by isolating the 80 kDa sperm protein by affinity chromatography. The present results suggest that the anti-80 kDa antibodies may be responsible for the infertility.


Phosphorylation of a 30,000 dalton protein during meitic maturation in Spisula oocytes


Protein phosphorylation during oocyte maturation in Asterias oocytes


Transblot identification of biotin-interacting proteins in rat liver


Protein phosphorylation during 5-hydroxytryptamine-induced maturation of Spisula oocytes

Maturation was induced in Spisula oocytes with 5-hydroxytryptamine (5-HT) creatinine sulfate at a final concentration of 5 microM. After 10 and 30 min of treatment, oocytes were homogenized and the cytosolic and particulate fractions were prepared. The fractions were incubated with [gamma-32P]GTP and [gamma-32P]ATP. The phosphorylated proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The radioactivity in the gels was determined by autoradiography. With [gamma-32P]GTP a marked increase in the radiolabeling of proteins with an estimated Mr of 47,000 and 20,000 in the cytosolic and particulate fractions, respectively, was demonstrated with the 5-HT-treated oocytes, whereas no stimulation was demonstrable with the use of [gamma-32P]ATP. A significant increase in GTP-mediated protein phosphorylation occurred within 10 min after 5-HT treatment before the occurrence of germinal vesicle breakdown, suggesting that this post-translation modification of proteins is an early action of the neurotransmitter in the induction of meiotic reinitiation in oocytes.


Identification of avidin-interacting proteins in Spisula oocytes: Correlation with germinal vesicle breakdown


Identification of cell types in Sertoli cell-enriched cultures by immunohistochemistry and DNA-specific fluorochrome Hoechst 33342

The cell types in Sertoli cell-enriched cultures can be identified by using the DNA-specific fluorochrome Hoechst 33342 staining. This simple, rapid and reproducible procedure can be used with fixed and living cells. The peritubular myoid cells can be distinguished from the Sertoli cells in Sertoli cell-enriched cultures by the characteristic staining pattern obtained using Hoechst 33342 dye. Those cells identified as peritubular myoid cells by the characteristic DNA staining also interacted with the anti-fibronectin antibody determined by an immunocytochemical method while the Sertoli cells did not. The described staining method is valuable in assessing the presence of peritubular myoid cells in Sertoli cell-enriched cultures.


The antibody from an infertile woman that induces sperm-agglutination interacts with rat testicular cells and sperm

The serum obtained from an infertile woman induced a specific head-to-head agglutination of human and rat sperm. The immunoglobulin G (IgG) fraction of the serum was obtained and found to interact with the proteins of rat sperm in testis and epididymis. Using an indirect immunofluorescent method with rat sperm from vas deferens, we determined that the antibody recognized the protein on the convex and concave regions of the acrosome and over the entire tail. However, with testicular spermatozoa, the antibody recognized only the distal end of the tails. In paraffin sections of adult rat testis, sperm tails located at the luminal region of the seminiferous tubules stained intensely. Weak but significant staining also occurred on late spermatids. In the epididymal sections, staining was restricted to spermatozoa in the lumen. On the other hand, sections of testes from 25-day-old rats containing spermatogonia and early spermatocytes had a completely negative reaction. Testicular somatic cells, including Sertoli cells, peritubular myoid cells and interstitial cells, did not stain. To identify the testicular protein interacting with the antibody, adult rat testis proteins were prepared and analyzed by a sodium dodecyl sulfate-polyacrylamide gel electrophoretic (SDS-PAGE) immunoblot technique. The antibody interacted with a protein with an estimated molecular weight of 82,000 in the testicular homogenate and particulate fraction, whereas the reaction was considerably weaker with the testicular cytosol fraction.


Identification of antigen in rat spermatogenetic cell interacting with an antihuman sperm monoclonal antibody

A monoclonal antibody (MAb) raised against human sperm protein, designated YWK-II, was used to determine the distribution of antigens in rat spermatozoa and rat testicular germ cells. By an indirect immunofluorescent method, the antibody localized over the rat spermatozoal head, except for the postacrosomal region. In paraffin sections of adult and immature rat testis, germ cells, at every developmental stage, and Sertoli cells stained, while interstitial cells and peritubular myoid cells remained unstained. When cocultures of Sertoli and germ cells were tested, only the germ cells stained intensely. Sertoli cells and peritubular myoid cells in cultures did not stain. In the epididymal sections, strong staining occurred with spermatozoa in the lumen and epididymal epithelial cells, with moderate staining in the myoid layers of epididymis. To determine the sperm antigen interacting with the YWK-II antibody, rat spermatozoa proteins were prepared and analyzed by an immunoblot technique. The monoclonal antibody interacted with a single protein, with an estimated molecular weight of 115,000, present in the cauda epididymal spermatozoa. Among the proteins of the caput epididymal spermatozoa, however, the antibody interacted with a major and a minor band with molecular weights of 115,000 and 88,000, respectively. On the other hand, with proteins prepared from the membrane fraction of adult and immature rat testis, the antibody reacted with two bands with estimated molecular weights of 88,000 and 115,000. In the lysate prepared from germ cells dissociated from Sertoli-germ cell cocultures, the antibody recognized only the 88,000 protein. The present results show that the YWK-II MAb interacts with two proteins with different molecular weights. The amount of the interacting proteins in spermatozoa varied with their location within the epididymis.


The Tokai Journal of Experimental and Clinical Medicine


Dibutyryl adenosine cyclic monophosphate regulates differentiation of type A spermatogonia with vitamin A in adult moue cryptorchid testes


Effects of duration of cryptorchidism in ability of mouse germ cells to regenerate and differentiate


Effects of Steel Mutation on Spermatogenesis Stimulated by Fetuin

The effect of the steel mutation on spermatogenesis was investigated using organ culture. Cultured cryptorchid testes from WB-+/+ mice showed an effective differentiation of type A spermatogonia in response to Pedersen type III fetuin. In contrast, the cryptorchid testes from WB-S1/+ showed neither differentiation nor division of type A spermatogonia. The findings reported here are the first demonstration of retarded response of steel mutation on a well-known agent, fetuin.


Vitamin A and Folicle-Stimulating Hormone synergistically induce differentiation of type A spermatogonia in adult mouse cryptorchid testis in vitro

To study the mechanism of vitamin A and FSH action on testicular germ cell differentiation, artificially induced cryptorchid testes of adult mice were cultured in vitro, because such testes contain only type A spermatogonia and Sertoli cells in their seminiferous tubules. A synergistic effect of vitamin A compounds with FSH, but not with LH, on testicular germ cell differentiation was observed. Our data suggested that FSH stimulated the proliferation of type A spermatogonia, whereas retinoids induce spermatogenesis from type A spermatogonia into intermediate or type B spermatogonia. Possible mechanisms of action of retinoids and FSH on testicular germ cell differentiation are discussed.


The effects of prostaglandin E2, parathyroid hormone, 1a, 25-dihydroxy-cholecalciferol and cyclic nucleotide analogs on alkaline phosphatase activity in osteoblastic cells

Clone MC3T3-E1 cells isolated from newborn mouse calvaria had the same type of alkaline phosphatase (ALP) as that found in adult mouse calvaria (the liver-bone-kidney type), as judged by polyacrylamide gel electrophoresis as well as by heat lability and amino acid inactivation. The effects of prostaglandin E2 (PGE2), parathyroid hormone (PTH), 1,25 dihydroxycholecalciferol [1,25(OH)2D3], and adenosine-3',5'-cyclic monophosphate (cAMP) analogs on ALP were investigated. PGE2 and 1,25(OH)2D3 increased ALP activity in dose-related manner with a maximal effect at concentrations of 10 ng/ml and 40 pg/ml, respectively. N6,O2-dibutyryl adenosine-3',5'-cyclic monophosphate (DBcAMP) also induced an increase in ALP activity in a dose-related fashion with a maximal effect at a concentration of 0.5 mM which was 2.2-fold over that of the controls. Induced ALP was of the "liver-bone-kidney" type. Antinomycin D and cycloheximide inhibited the increase in ALP activity induced by DBcAMP. The level of ALP was elevated by 8-bromo-adenosine-3',5'-cyclic monophosphate and theophylline, but not by N6,O2-dibutyryl guanosine-3',5'-cyclic monophosphate and sodium butyrate. Moreover, PGE2 dramatically increased the level of cAMP in the cells with a maximal effect at a concentration of 10 ng/ml, indicating that PGE2 and DBcAMP induce an increase of ALP activity in clone MC3T3-E1 cells by increasing the cAMP level; PTH did not affect enzyme activity and cAMP level in the cells.(ABSTRACT TRUNCATED AT 250 WORDS)


In vitro demonstration of deleterious effect of steel mutation on spermatogenesis in mice

The effect of the steel mutation on spermatogenesis was investigated by using organ culture. Cultured cryptorchid testes from C57BL/6- +/+ mice showed an effective differentiation of type A spermatogonia in response to an increased concentration of bovine serum. In contrast, the cryptorchid testes from C57BL/6- Sld/+ mice showed only limited differentiation of type A spermatogonia even in the highest concentration of serum tested. The type A spermatogonia of mutant mice showed the normal mitotic response but did not differentiate further. The findings reported here are the first in vitro demonstration of the deleterious effects of the steel mutation on testicular germ cell differentiation and suggest that the effect of the steel mutation is expressed in Sertoli cells, the only supporting element in seminiferous tubules, or germ cells themselves.


1α, 25-dihydroxyvitamin D3 and analogues of vitamin D3 induce alkaline phosphatase activity in osteoblastic cells derived from newborn mouse calvaria

In order to study the effects of vitamin D metabolites on bone metabolism, clone MC3T3-E1 cells, which have retained osteoblastic activity, were cultured with various concentrations of the hormone, 1 alpha, 25-dihydroxyvitamin D3 [1 alpha, 25 (OH)2D3]. A physiological concentration of 1 alpha, 25 (OH)2D3 stimulated alkaline phosphatase (ALP) activity in the cells. Other metabolites--1 alpha, 24-dihydroxyvitamin D3 [1 alpha, 24 (OH)2D3], 1 alpha-hydroxyvitamin D3 [1 alpha (OH)D3], and 24R,25-dihydroxyvitamin D3 [24R,25 (OH)2D3]--also induced increases in ALP activity in a dose-dependent fashion. However, their effective concentrations were 100 or 1,000 times greater than that of 1 alpha, 25 (OH)2D3. Hormone-induced and native ALP activities in the cells were of the same type as that found in newborn mouse calvaria; that is, they were heat-labile, L-homoarginine- and levamisole-sensitive, and L-phenylalanine-insensitive (liver-bone-kidney type). These results show that vitamin D metabolites stimulate bone formation in vitro and that they may be involved in bone formation in vivo as well.


Retinoids induce differentiation of type A spermatogonia in vitro: organ culture of mouse cryptorchid testes.

To study the effects of retinoids on testicular germ cell differentiation, especially on stem cells, artificially induced cryptorchid testes of adult mice were cultured in vitro inasmuch as they contain only stem cells, type A spermatogonia. We report here that retinol, retinal, retinol acetate, even retinoic acid, activated cell division in type A spermatogonia and stimulated them to differentiate. These findings are the first demonstration of the in vitro induction of early stage of spermatogenesis by retinoids and also retinoic acid, which is considered to be a biochemically inactive compound for mammalian testicular function.


In vitro differentiation of type A spermatogonia from mouse cryptorchid testis in serum-free media

Artificially induced cryptorchid testes of adult mice were cultured in serum-free media. Pedersen Type III fetuin activated cell division in Type A spermatogonia and stimulated them to differentiate; it also had a synergistic effect with follicle-stimulating hormone (FSH) on testicular germ cell differentiation. However, Pedersen Type IV, Deutsch, and Spiro fetuin did not show any stimulatory effect on the differentiation of germ cells. It was suggested that some serum factor(s) which copurifies with Pedersen Type III fetuin may be the active principle rather than fetuin itself in the differentiation of testicular germ cells in vitro.


Hormones and the differentiation of type A spermatogonia in mouse cryptoorchid testis incubated in vitro

In order to study hormonal effects on testicular germ cell differentiation, especially on type A spermatogonia, artificially induced cryptorchid testes of adult mice were cultured in a medium containing testosterone, dihydrotestosterone, tri-iodothyronine, dibutyryl 3':5' cyclic adenosine monophosphate, human chorionic gonadotrophin, LH, FSH, insulin and transferrin. These substances, with the exception of FSH, showed no stimulatory effect on the differentiation of type A spermatogonia. However, FSH activated cell division in type A spermatogonia and stimulated them to differentiate, while LH showed neither the promotion of differentiation nor a synergistic effect on FSH-mediated germ cell differentiation.


Retinoic acid (Vitamin A acid) induces spermatogenesis in adult mouse cryptorchid testis in vitro

Biochemical and Biophysical Research Communications


Testicular DNA synthesis in vivo: Changes in DNA synthetic activity following artificial cryptorchidism and its surgical reversal


Testicular DNA synthesis in vivo: Comparison between unilaterally cryptorchid testis and contralateral intact testis in mouse

DNA synthetic activity was studied by autoradiography in unilaterally cryptorchid (abdominal) and contralateral intact(scrotal) testes in mice. Nongerm cells were inactive in DNA synthesis in both the cryptorchid testis and in the intact control. Labeling indices of undifferentiated type A spermatogonia did not show any differences, indicating that type A spermatogonia did not temperature sensitive in DNA synthesis in vivo.


Effects of hormones on the differentiation of type A spermatogonia in mouse cryptorchid testis in vitro

実験的マウス停留睾丸におけるタイプA精原細胞の分化に及ぼすホルモンの影響について


Differential effects of cyclic AMP on DNA synthesis by germ cells and non-germ cells in mouse testicular culture


The effects of steel mutation on testicular germ cell differentiation

The effects of artificial cryptorchidism and its surgical reversal on spermatogenesis were examined in germ cell mutant, S1/+ and wild type, +/+, mice. In cryptorchid testes no difference was found between S1/+ and +/+ mice in the number of undifferentiated type A spermatogonia. The activity of type A spermatogonia in mutant mice appeared normal as judged by its mitotic cell number and DNA synthesis. The surgical reversal of cryptorchidism resulted in regenerative differentiation of mature germ cells in both types of mice, but the pattern of cellular differentiation in the mutant testes was completely different from that of the wild type testes. At two steps of cellular differentiation, intermediate or type B spermatogonia and spermatid, the numbers of cells were much smaller in the S1/+ testes than those in the +/+ testes. The steel gene was therefore suggested to exert its effects on the differentiation of type A spermatogonia to intermediate or type B spermatogonia, on meiotic division and/or the survival rate of these cells, but not on the undifferentiated type A spermatogonia or stem cells.