• 제목/요약/키워드: adult testicular cells

검색결과 30건 처리시간 0.031초

Beta-carotene prevents the spermatogenic disorders induced by exogenous scrotal hyperthermia through modulations of oxidative stress, apoptosis, and androgen biosynthesis in mice

  • Yon, Jung-Min;Kim, Jae Seung;Lin, Chunmei;Park, Seul Gi;Gwon, Lee Wha;Lee, Jong-Geol;Baek, In-Jeoung;Nahm, Sang-Seop;Nam, Sang-Yoon
    • 대한수의학회지
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    • 제59권2호
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    • pp.59-67
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    • 2019
  • We investigated whether ${\beta}$-carotene (${\beta}-CA$) or ellagic acid (EA), originating from various fruits and vegetables, has a preventive effect against male infertility induced by exogenous scrotal hyperthermia. ICR adult mice were intraperitoneally treated with 10 mg/kg of ${\beta}-CA$ or EA daily for 13 days consecutively. During this time, mice were subjected to transient scrotal heat stress in a water bath at $43^{\circ}C$ for 20 min on day 7, and their testes and blood were obtained on day 14 for histopathologic and biochemical analyses. Heat stress induced significant testicular weight reduction, germ cell loss and degeneration, as well as abnormal localization of phospholipid hydroperoxide glutathione peroxidase (PHGPx) and manganese superoxide dismutase (MnSOD) in spermatogenic and Leydig cells. Heat stress also altered the levels of oxidative stress (lipid peroxidation, SOD activity, and PHGPx, MnSOD, and $HIF-1{\alpha}$ mRNAs), apoptosis (Bax, Bcl-xL, caspase 3, $NF-{\kappa}B$, and $TGF-{\beta}1$ mRNAs), and androgen biosynthesis (serological testosterone concentration and $3{\beta}$-hydroxysteroid dehydrogenase mRNA) in testes. These changes were all improved significantly by ${\beta}-CA$ treatment, but only slightly improved by EA treatment. These findings indicate that ${\beta}-CA$, through modulations of oxidative stress, apoptosis, and androgen biosynthesis, is a potent preventive agent against testicular injuries induced by scrotal hyperthermia.

Glucocorticoids improve sperm performance in physiological and pathological conditions: their role in sperm fight/flight response

  • Vittoria Rago;Adele Vivacqua;Saveria Aquila
    • Anatomy and Cell Biology
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    • 제57권1호
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    • pp.119-128
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    • 2024
  • Glucocorticoids play a physiologic role in the adult male reproductive functions, modulating gonadal steroid synthesis and spermatogenesis, through the glucocorticoid receptor (GR). The expression of GR has been described in several key testicular cell types, including somatic cells and early germ cell populations. Nothing is known on GR in human spermatozoa. Herein, we explored the GR expression and its possible role in normal and testicular varicocele semen samples from volunteer donors. After semen parameter evaluation by macro- and microscopic analysis, samples were centrifuged; then spermatozoa and culture media were recovered for further investigations. By western blotting and immunofluorescence analyses we evidenced for the first time in spermatozoa the presence of GR-D3 isoform which was reduced in sperm from varicocele patients. By treating sperm with the synthetic glucocorticoid dexamethasone (DEXA), we found that survival, motility, capacitation, and acrosome reaction were increased in both healthy and varicocele samples. GR involvement in mediating DEXA effects, was confirmed by using the GR inhibitor mifepristone (M2F). Worthy, we also discovered that sperm secretes different cortisol amounts depending on its physio-pathological status, suggesting a defence mechanism to escape the immune system attach in the female genital tract thus maintaining the immune-privilege as in the testis. Collectively, our data suggests a role for glucocorticoids in determining semen quality and function, as well as in participating on sperm immune defensive mechanisms. The novelty of this study may be beneficial and needs to take into account in artificial insemination/drug discovery aimed to enhancing sperm quality.

Expression of Cyclin D3 Transcripts in the Postmeiotic Male Germ Cells of the Mouse

  • Sun, Woong-Sun;Geum, Dong-Ho;Choi, Wan-Sung;Kim Kwon, Yun-Hee;Rhee, Kun-Soo;Kim, Kyung-Jin
    • Animal cells and systems
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    • 제2권4호
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    • pp.495-500
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    • 1998
  • D-type G1 cyclins are known to be crucial for the progression of mitotic cell cycle in mammals. Although many studies have been performed to elucidate the roles of D-type cyclins, it is largely unknown whether D-type cyclins are directly involved in the regulation of meiotic germ cell development. In the present study, we examined the expression patterns of D-type cyclins (cyclin D1 and D3) during male germ cell development by northern blot and in situ Hybridization analyses. In the adult testes, we detected a 4.2 kb cyclin D1 mRNA and two different sizes (2.3 kb and 1.8 kb) of cyclinD3 mRNAs. The short form of the cyclin D3 transcript was testis-specific. Along with the testicular development, expression of cyclin D3 mRNA was increased whereas cyclin D1 mRNA was gradually decreased. in situ hybridization study also revealed that the expression of cyclin D3 was restricted to the postmeiotic germ cells. Furthermore, the 2.3 kb transcript was highly expressed in the round spermatids and decreased in the elongated spermatids/residual bodies, while the 1.8 kb transcript was expressed in elongated spermatids/residual bodies more abundantly. Sucrose-gradient separation of polysomal RNA fractions demonstrated that some portions of the 2.3 kb transcript are translationally active, while the 1.8 kb transcript is likely to be inactive. Taken together, the present data suggest a functional importance of cyclin D3 expression in the differentiated postmeiotic male germ cells.

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토끼에서 출생 후 고환간질세포의 발생에 관한 연구 (Studies on the postnatal development of the Leydig cell in rabbits)

  • 태현진;박영재;강형섭;김남수;박상열;양홍현;안동춘;김인식
    • 대한수의학회지
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    • 제45권3호
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    • pp.325-334
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    • 2005
  • Changes in the rabbit Leydig cell from birth to adulthood were studied in New Zealand white rabbits of 1, 7, 21, 35, 49, 70, 105, 147, 196, and 252 days (n = 8 rabbits per group) of age. The objectives of this study were to understand the fate of the fetal Leydig cells, to determine the changes in serum testosterone levels, and leutenizing hormone-stimulated testosterone production per testis in vitro, and to quantify adult Leydig cells by number and average volume with age. Testes of rabbits were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative (stereological) morphological studies were performed. Testosterone levels in the incubation medium of luteinizing hormone-stimulated (100 ng/ml) testosterone secretion per testis in vitro, and in serum were determined by radioimmunoassay. The average volume of a testis of 1-day-old rabbits was determined as $0.0073cm^3$ and the parameter increased linearly from birth to 252 days ($3.93cm^3$). The volume density of the seminiferous tubules increased with age from 33.76% at day 1 to 88.2% at day 252. The volume density of the interstitium represents 66.24% of the testicular parenchyma at day 1. This proportion progressively diminished during development to reach a value of 11.8% at day 252. The volume density of Leydig cells increased almost linearly from birth (0.001%) to 252 days (2.62%). Leydig cell mass per testis increases from 0.0012 mg to 0.25 mg between days 1 and 35, from 2.66 mg to 44.3 mg between days 49 and 105 and from 65.42 mg and 102.9 mg between days 147 and 252. The absolute numbers of adult Leydig cells per testis increased linearly from birth to 252 days. The average volume of adult Leydig cell on days 1, 7, 21 and 35 was not significantly different; a gradual and continued increase was observed thereafter, reaching a 3-fold increase at 196 and 252 days. Serum testosterone concentrations were not significantly different at day 1 compared days 7, 21, 35. Significant increases were observed at days 49 and 70. Values at days 70 and 105 and days 147, 196, and 252 were not significantly different. LH-stimulated testosterone production per testis in vitro was significantly different at day 1 compared days 7, 21, 35. Significant increases were observed at days 49 and 70. Hormonal values at days 105, 147, 196, and 252 were not significantly different. These data suggested Leydig cell developmental phase can be classified: a neonatal phase (1-7 days), a prepubertal phase (14-49 days) and an adult phase (70-252 days). Immature and mature adult Leydig cells, initially detected at days 7 and 49, respectively, and mature adult Leydig cells were abundant Leydig cell type according to the number and absolute volume per testis form day 49 onwards.

Dietary crude protein levels during growth phase affects reproductive characteristics but not reproductive efficiency of adult male Japanese quails

  • Retes, Pamela Lacombe;Neves, Danusa Gebin das;Bernardes, Laryssa Fernanda;Alves, Victoria Veiga;Goncalves, Natalia de Castro;Lima, Diego de Rezende;Alvarenga, Renata Ribeiro;Pereira, Barbara Azevedo;Seidavi, Alireza;Zangeronimo, Marcio Gilberto
    • Animal Bioscience
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    • 제35권3호
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    • pp.385-398
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    • 2022
  • Objective: The objective was to evaluate the influence of different dietary crude protein (CP) levels during the growth phase on reproductive characteristics and reproductive efficiency as well as the body development of adult male Japanese quail. Methods: Three hundred one-day-old male quails were distributed into five treatments with diets containing different CP levels (18%, 20%, 22%, 24%, and 26%) in a completely randomized design, with six replicates of ten birds each. The CP diets were applied only during the growth phase (1 to 35 days). At 36 days of age, the birds were transferred to 30 laying cages with three males and nine females each, and all birds received the same diet formulated to meet production-phase requirements until 96 days of age. Results: The growth rate of the birds increased linearly (p<0.01) with increasing dietary CP, but the age of maximum growth decreased (p<0.05). At growth maturity, all birds had the same body weight (p>0.05). At 35 days of age, higher weight gain was obtained (p<0.05) with diets containing 22% CP or higher. No effects on feed conversion were observed in this phase. The increase in dietary CP enhanced (p<0.01) nitrogen intake and nitrogen excretion but did not affect (p>0.05) nitrogen retention. Testis size, seminiferous tubular area, number of spermatogonia, and germinal epithelial height at 35 days of age increased linearly (p<0.05) with dietary CP, while the number of Leydig cells decreased (p<0.01). The Sertoli cell number at 60 days of age increased linearly (p<0.01) with dietary CP. Dietary CP levels did not affect cloacal gland size, foam weight, foam protein concentration, semen volume, or flock fertility at 90 days of age. Conclusion: Dietary CP concentration affected body and testicular development in male Japanese quails but did not affect reproductive efficiency.

EST Clustering 방법으로 동정한 새로운 유전자의 생쥐 난소 및 정소에서의 발현 (Identification of a Novel Gene by EST Clustering and its Expression in Mouse Ovary and Testis)

  • 황상준;박창은;황규찬;이경아
    • Clinical and Experimental Reproductive Medicine
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    • 제33권4호
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    • pp.253-263
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    • 2006
  • 목 적: 본 연구에서는 EST Clustering 방법을 이용하여 이전의 연구에서 발굴한 B357 EST의 염기서열을 포함하는 유전자를 동정하고, 이 유전자의 발현을 생쥐의 난소와 정소를 포함한 여러 가지 조직들에서 살펴보고자 하였다. 연구방법: EST Clustering으로 얻어진 전체 염기서열을 5-day-ovary-specific gene-1 (5DOS1)이라고 명명하여 GenBank에 등록하였으며 (AY751521), northern blotting, real-time RT-PCR, in situ hybridization, western blotting, immunohistochemistry 등의 방법을 이용하여 생쥐 난소와 고환의 발달단계에 따라 그 발현양상을 관찰하였다. 결 과: 5DOS1의 전사체는 성장한 정소, 뇌, 근육에서 높게 발현하였으며, 난소의 경우에서는 원시난포시기부터 모든 난자에서 발현하였으며 특히 생후 5일째 높게 발현하였고 그 이후로는 점차 감소하였다. 반면 정소의 경우는 발달과 함께 계속 증가하였으며, 정모세포를 제외한 모든 발달단계별 정자에서 발현함을 관찰하였다. 결 론: 본 연구결과는 5DOS1에 대한 발견과 동정에 대한 첫 보고로써, 유전자 및 단백질이 생쥐의 난소 및 정소의 생식세포에서 발현하는 것을 관찰하였다. 앞으로 생식세포 발생 및 분화에 관련된 5DOS1의 기능에 대한 심층 연구가 더 필요하다고 사료된다.

한국 재래 닭 고환 간질세포의 미세구조에 관한 연구 (The Ultrastructure of Leydig Cells in the Testis of Korean Native Chickens)

  • 장병귀;태현진;최철환;박영재;양홍현;김남수;박상열;강형섭;박병용;이영훈;안동춘;김인식
    • 한국가금학회지
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    • 제33권3호
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    • pp.181-188
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    • 2006
  • 한국 재래 닭에서 부화 후부터 성숙에 이르는 시기까지 고환 발달동안 간질세포의 미세구조를 구명하기 위하여 부화 후 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 28, 32, 44, 52 및 64주령(n=13마리/일령)의 한국 재래 닭을 이용하여 이 연구를 수행하였다. 한국 재래닭의 고환은 2.5% glutaraldehyde를 이용하여 전신관류고정하고 조직 처리 과정을 거쳐 Epon-araldite에 포매하였다. 초박절편기를 사용하여 $1{\mu}m$로 절편한 다음 methylene blue로 염색하여 일반적인 조직의 변화상을 관찰하였고 간질세포의 미세구조는 초박 절편을 제작하여 투과전자현미경으로 관찰하였다. 간질세포내에서 테스토스테론과 연관성이 있는 미세구조인 사립체, 평활형질내세망 및 지방방울의 변화에 따라 간질세포의 발달 유형을 분류하면, 1) 간질세포가 치밀하게 배열되어 있고 세포 사이의 공간이 거의 없으며 핵이 간질세포 용적의 많은 부분을 차지하고 있는 시기로 핵은 전자밀도가 높은 유형과 희박한 유형으로 나눌 수 있었고, 또한 세포질 내에는 많은 양의 지방방울, 소량의 사립체 및 평활형질내세망을 갖고 있는 시기, 2) 세포질 내에 큰 폭으로 확장된 평활형질내세망, 풍부한 사립체 및 감소된 지방방울을 갖고 있는 시기로 나눌 수 있다.

Intratesticular Injection of Hypertonic Saline: Non-Invasive Alternative Method for Animal Castration Model

  • Kwak, Byung Kuk;Lee, Sung-Ho
    • 한국발생생물학회지:발생과생식
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    • 제17권4호
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    • pp.435-440
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    • 2013
  • Previous studies, including our own, have demonstrated that the intratesticular injection of hypertonic saline (20%) decreased serum testosterone level which was similar to the surgical castration in the rat, showing the state of chemical castration. In the present study, we further verify the efficacy of this less invasive method as an alternative of surgical orchidectomy in the andrological field. Sterilized 20% saline was directly injected into the adult male rats (750 ${\mu}l$ per testis). The tested rats were divided into 3 groups including intact group (intact), orchidectomy group (ORX) and saline injection group (SAL) after bilateral orchidectomy was performed at the same day of injection. All rats were sacrificed at 4 weeks after injection. The reproductive organs (testes, epididymis, seminal vesicles and prostates) were collected and used for DNA and protein pattern analyses. Also, patho-histological studies on the testes were performed. In contrast to the intact group, similar DNA damages of testis and seminal vesicle were appeared in ORX group and SAL group. The DNA degradations seemed to be the results of necrosis rather than apoptosis. In the protein pattern analysis, all the testing tissues exerted similar patterns in the ORX group and the SAL group compared to the those of intact group. Patho-histological studies revealed that severe degenerative changes in testicular seminiferous tubules and massive infiltration of immune cells in SAL group. The present study confirmed that direct injection of hypertonic saline into the testis caused the equivalent biochemical changes in the accessory sex organs as shown in the orchidectomized animals. These results suggest that hypertonic saline injection model could be a useful castration model which can substitute for surgical castration when its safety is secured through further study in the future.

고장성 식염수 주사에 의한 흰쥐 정소의 해부학적 변화 (Histological Changes in Rat Testis by Injection of Hypertonic Saline)

  • 곽병국;이철상;이성호
    • 한국발생생물학회지:발생과생식
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    • 제14권4호
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    • pp.281-286
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    • 2010
  • 최근 고장성(20%) 생리식염수를 흰쥐 정소에 직접 주사할 경우, 혈중 테스토스테론 수준이 최저 수준에 도달하는 화학적 거세 상태에 들어감이 보고되었다. 본 연구는 이러한 간단한 생리 식염수 주사 모델의 효용성을 더 검증하기 위해 정소의 미세 해부 구조상의 변화를 조사하였다. 실험군으로는 무처리군(intact, control), 정소제거군(orchidectomy), 그리고 식염수 주사군(saline-injection)으로 하였으며, 식염수 주사군에는 정소당 $750{\mu}{\ell}$의 멸균한 20% 식염수를 직접 주사하였다. 정소 제거 수술은 식염수 주사가 행해진 날에 시행하였다. 주사후 30일 경과 후에 동물을 희생하여 생식조직들을 채취하고 무게를 측정한 후 조직학적 조사를 위해 고정시켰다. 체중의 경우 무처리 대조군과 비교하여 정소절제군과 식염수 주사군 모두에서 유의한 변화는 없었다. 식염수 주사군의 정소 무게는 무처리 대조군에 비해 유의하게 감소하였다. 부정소와 저정낭의 무게의 경우 무처리 대조군에 비해 정소절제군과 식염수 주사군 모두에서 유의하게 감소하였으며, 전립선의 경우 무처리 대조군에 비해 정소절제군에서는 유의하게 감소하였으며, 식염수 주사군에서는 감소하는 경향은 있었지만 유의성은 없었다. 외양상으로 식염수 주사군의 정소에는 약간의 위축이 나타났으며, 백막이 정상적으로 보였다. 그러나 조직학적으로 정소 대부분에서 괴사가 발견되었으며, 헤마톡실린-에오신으로 거의 염색되지 않았다. 동일한 절편에서, 주사 부위의 정반대 부분은 비정상적인 세포층들이 염색되었다. 대부분의 세정관에서 미성숙한 생식세포들이 기질 막에서 분리되고, 내강 쪽으로 세포들이 떨어져 나간 것이 확인되었다. 본 연구는 정소내 고장성 식염수 직접 주사가 부속 생식기관에 정소절제와 유사한 테스토스테론 제거 효과를 유발할 수 있음을 확인하였다. 비록 심층적인 연구들이 더 요구되지만, 본 연구는 정소 내 고장성 식염수 직접 주사법이 비용이 덜 들고 덜 침습적이면서 정소절제술이나 화학적 거세와 거의 동일한 효용성을 가졌음을 시사한다.

태생 경골어류, 망상어(Ditrema temmincki)의 초기생식소 형성 및 성분화 (Early Gonadogenesis and Sex Differentiation in the Viviparous Teleost, Ditrema temmincki)

  • 이정식;이영돈
    • 한국수산과학회지
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    • 제29권1호
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    • pp.35-43
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    • 1996
  • 망상어, Ditrema temmincki이 시원생식세포의 출현, 원시생식소와 초기생식소의 형성, 정소와 난소의 분화 및 체내자어의 성비조성을 조직학적으로 조사하였다. 시원생식세포는 부화전 배의 초기소화관과 등쪽 체벽사이의 섬유성 간충직에서 처음 식별되었다. 부화후부터 전장 4.0mm 까지 소화관과 체벽사이의 섬유성 간충직에 고르게 분포하던 시원생식세포들은 간충직이 등쪽 체벽으로 부터 분리되는 전장 5.0 mm 시기를 전후해서 간충직의 후방으로 이동하여 원시생식소를 형성하게 된다. 초기생식소 형성과정 동안 정소와 난소의 분화는 생식세포와 체세포의 배열 상태에 의하여 구분되며, 전장 10.0mm 부터 생식소는 외부형태에 의하여 정소와 난소로 구분되는데, 정소는 한쌍으로 분리된 형태이고 난소는 후반부가 융합된 형태이다. 정소의 분화는 체내자어의 크기가 전장 25.0mm를 전후해서 정소에 곡정세관의 조직상이 나타나고, 전장 30.0mm를 전후해서 수정관이 형성되며, 전장 45.0 mm를 전후해서 외부형태와 내부 구조적으로 성어와 유사한 정소로 분화된다. 난소의 분화는 체내자어의 크기가 전장 30.0mm를 전후해서 난소습곡과 난소강의 형성이 명확하며, 전장 60.0mm를 전후해서 외부형태와 내부 구조적으로 성어와 유사한 난소로 분화된다. 체내자어의 출산시 전장은 63.0mm 전후이고, 이때 정소와 난소에는 각각 정원세포와 염색인기 단계의 생식세포를 보유하며, 암 : 수 성비는 약 1.65 : 1이었다. 망상어의 성은 자웅이체이며, 성분화 양식은 분화형에 속한다.

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