Inverse relationship between testicular proliferation and apoptosis in mammalian seasonal breeders

Seasonal cycles of testicular activity occur in many mammals and can include transitions between total arrest and recrudescence of spermatogenesis. We hypothesize that involution and reactivation of testis result from two antagonistic processes, proliferation and programmed cell death (apoptosis), which are activated at different times. To test this hypothesis, quantitative measurements of both proliferation-specific marker and apoptotic produced nucleosomes have been compared with sperm and testosterone production in testes from adult roe deer during breeding and non-breeding seasons (May to September). Testes of brown hare were included from periods of testes regression (June to August) and recrudescence (November to December). The highest testicular weights in roe deer were found in the rutting period from late July to early August (27.25 +/- 8.56 g), corresponding with the highest number of testicular sperm/g parenchyma. The peak of sperm production coincided with a peak in testosterone concentration (1.19 +/- 0.53 microg/g testis). The maximum level of proliferation-specific marker was also found during the breeding season (98.6 +/- 58.2 U/g testis in comparison to 20.1 +/- 22.0 U/g in the prerutting period). In contrast, the most significant apoptosis was observed in the nonbreeding season than the breeding period (71.11 +/- 5.79 U/mg testis and 18.88 +/- 6.79 U/mg, respectively). Testicular proliferation was low in the brown hare (0.061 +/- 0.062 U/g) during involution of the testes. It was newly activated in November and December (0.85 +/- 0.33 U/g), preceding the increase in testicular volume. Testosterone production increased in conjunction with testicular proliferation. At this time, testicular apoptosis was significantly lower (14.16 +/- 2.12 U/mg testis) than during the period of pronounced testicular regression (30.16 +/- 19.95 U/g). These results suggest that regulation of seasonal testicular activity is characterized by an inverse relationship of proliferation and apoptosis.     

Reproductive pattern in the male Angolan free-tailed bat, Tadarida (Mops) condylura (Microchiroptera: Molossidae) in the Eastern Transvaal, South Africa

The reproductive pattern in male Angolan free-tailed bats Tadarida (Mops) condylura was studied at two localities in the Eastern Transvaal during the period 1989-1991. Males displayed a nine-month breeding season extending from June to February. Following testicular recrudescence in early June, spermatogenic activity displayed two periods of peak activity in August/early September and November/early December. Although testes became involuted in March, baseline spermatogenic activity was maintained and tests never regressed to prepubertal conditions. Leydig cell morphology closely followed spermatogenic activity, with the diameter of Leydig cell nuclei displaying a concomitant seasonal pattern.     

Seasonally activated spermatogenesis is correlated with increased testicular production of testosterone and epidermal growth factor in mink (Mustela vison)

Seasonal changes in spermatogenesis were studied with respect to testicular production of both testosterone and epidermal growth factor (EGF) in mink. The testes were collected in November (n = 15; testis recrudescence), February (n = 15; before breeding season), March (n = 14; breeding season), and May (n = 11; testis involution) and the following parameters of testicular activity were quantified: testicular mass, number of testicular spermatozoa, percentages of haploid, diploid, and tetraploid (G2/M-phase) cells and content of testosterone and EGF. The growth factor was immunohistochemically localized in the parenchyma. Testis mass, spermatogenic activity, and the production of both testosterone and EGF were maximal in March, but were not significantly different from the levels in February. The correlation between testis weight and sperm per testis was r = 0.825 (P < 0.001). Testosterone and EGF levels were correlated to each other (r = 0.78; P < 0.001) and had significant positive correlations to testis mass, number of sperm and proportion of haploid cells; and negative correlations to percentages of mitotic cells. EGF was localized in interstitial cells and in the luminal region of seminiferous tubules, where it occurred during the last steps of spermiogenesis. We inferred that intensified seasonal spermatogenesis was stimulated by testosterone and by autocrine/paracrine effects of EGF.     

Seasonal patterns of ornithine decarboxylase activity and levels of polyamines in relation to the cytology of germinal cells during spermatogenesis in the sea star, Asterias vulgaris

The activity of ornithine decarboxylase (ODC) and levels of polyamines were measured in the testes of Asterias vulgaris collected throughout an annual spermatogenic cycle. Samples of the testes were prepared for light and electron microscopy to observe the associated changes in the cytology of germinal cells. The specific activity of ODC increased at the onset of testicular growth, decreased during the coldest period of the winter when testicular growth was minimal, and increased again early in the spring when testes grew maximally. Increased activity of ODC resulted in increased levels of polyamines and was correlated with either mitotic or meiotic germinal cell divisions, or both. Spermine levels were always greater than putrescine, followed by spermidine. Highest levels of polyamine synthesis coincided with the onset of spermatogonial proliferation during the fall and with the period of meiotic differentiation and spermiogenesis in the spring. Mid-summer (July) testes were small (0.3-0.5 gonad index (GI)) and contained amitotic spermatogonia arrested in G(1) of the cell cycle. Mitotic and pre-meiotic testes (October/November) increased slightly in size (0.3-1.4 GI) and contained actively dividing spermatogonia, most of which differentiated into primary spermatocytes. Testes from February and March were large (1-6.75 GI), but the proliferative status of their spermatogonia and primary spermatocytes varied. Spermatogonia and primary spermatocytes from early February testes were not dividing. In testes obtained in March, both spermatogonial mitosis and meiosis of spermatocytes resumed, coincident with increased field water temperatures.     

Testicular response to melatonin or suprachiasmatic nuclei ablation in the spotted skunk

Testes of the Western spotted skunk enlarge and regress seasonally. The pineal hormone, melatonin, may be important in timing this seasonal reproductive activity. Likewise, the suprachiasmatic nuclei (SCN) have been implicated as possible neural regulators of pineal and reproductive events. These experiments were conducted to determine whether ablation of the SCN or constant administration of melatonin alters timing of the seasonal pattern of testicular regression and recrudescence. Male skunks (n = 24) were treated as follows: six received two empty Silastic capsules, six received two melatonin-filled Silastic capsules, six received sham lesions to the SCN, and six received lesions to the SCN (SCNx). All skunks were exposed to a natural photoperiod and had regressed testes at the onset of the experiment. Four of six males from the SCNx group had an average of 94 +/- 11.3% of these nuclei destroyed. Sham SCNx, animals with less than 40% of the SCN ablated, and males with empty capsules did not have fully enlarged testes until October. SCNx and melatonin-treated skunks exhibited a hastening of testicular recrudescence with maximal testis size being reached in June. Skunks with lesions to the SCN maintained enlarged testes for 5 months while all other groups exhibited rapid regression after attaining maximal testis size. Testicular regression occurred from July through October in animals receiving continuous melatonin, while controls exhibited recrudescence during this same period. Our data suggest that the SCN, melatonin, and thus the pineal gland, play a role in regulating the seasonal testicular cycle of the spotted skunk.     

Spermatogenesis, testicular composition and the concentration of testosterone in the equine testis as influenced by season

The influence of season on spermatogenesis, testicular composition and the concentration of testosterone in the equine testis was evaluated using testes from 45 stallions. Testes were obtained through a commercial abbatoir during September, December-January, March and July. The weights of the testes, the tunica albuginea and testicular parenchyma and the proportion of the testicular parenchyma occupied by seminiferous tubules or interstitial tissue were similar during each season. How ever, diameter of the seminiferous tubules was greater in July than during other months of the study. In addition, the concentration of testosterone within the testicular parenchyma was twice as great during July as during the fall and winter, and this period of peak testicular testosterone concentrations was associated with spermatozoal production rates, which were 65% greater than those observed in September.     

Annual changes in testicular development and occurrence of parasperm in the male reproductive organs of fourspine sculpin, Cottus kazika

Annual changes in testicular development and occurrence of parasperm were investigated using 2-year-old male fourspine sculpins Cottus kazika, based on the histological observation of testes. The male reproductive organ of fourspine sculpins comprised a pair of testes and a sperm duct that functioned as a sperm-storage organ. Male maturity was divided into the following periods: spermatogonial proliferation period (September), early spermatogenic period (October), mid-spermatogenic period (November), late spermatogenic period (December and January), functional maturation period (February and March), and recovery period (April to August). Spermatogenesis rapidly progressed from October to January and continued until the functional maturation period. Parasperm formation, which is known in some cottidae species, was observed in fourspine sculpins. Testicular regression of cultured fourspine sculpins progressed slowly during the recovery period when residual parasperm and empty spaces occupied the testis. The parasperm were immotile and oval and slightly concave on one side; additionally, they stained strongly with hematoxylin and PAS. Seminal lobules of the testis were filled with parasperm during the spawning period; in contrast, the sperm duct was filled with eusperm. These findings were observed in both cultured and wild fish. In this study, the functions of parasperm with regard to reproduction in fourspine sculpins are discussed.     

Seasonal reproduction of a tropical bat, Anoura geoffroyi, in relation to photoperiod.

Anoura geoffroyi (Chiroptera, Phyllostomidae, Glossophaginae), Geoffroy's hairy-legged long-tongued bat, were collected from September 1984 to August 1985, and these bats were found to breed seasonally in the wild on Trinidad, West Indies, at 10 degrees N latitude. Histological examination of these samples indicated that females became pregnant in July or August, and young were born in late November or early December. The testes and epididymides were small from September to mid-April, increased threefold in weight between mid-April and late May, reached a peak weight in July, and decreased in weight in August. Spermatogenesis occurred throughout the testes of males captured from May to August. In 1990, the timing of parturition in females that gave birth in the laboratory to young conceived in the wild was similar to the timing in the field in 1984-1985. Groups of 10-13 males were subjected in the laboratory to (i) a gradually changing, civil twilight photoperiod that mimicked the natural cycle of annual change at 10 degrees N latitude, (ii) the same gradually changing cycle of photoperiod accelerated to a six-month period, or (iii) a constant photoperiod (light 12:54 h: dark 11:06 h). These treatments began in mid-December, four months before the initiation of testicular recrudescence in the wild. In all three groups, testicular volume remained low until April, and then increased two- to threefold between late April and late June, rising to a peak in July, as occurred in the wild.(ABSTRACT TRUNCATED AT 250 WORDS)     

The annual testicular cycle in an equatorial colony of lesser rock hyrax, Heterohyrax brucei.

Adult males from a colony of lesser rock hyrax found near the equator in Kenya exhibited an annual cycle of testicular activity characterized by intense spermatogenesis and elevated androgen status from May to July. Average masses of testes and seminal vesicles taken in these months were almost fourfold greater than those from September to January. During the months of peak testicular activity average diameters of Leydig cells and seminiferous tubules were increased by approximately one half and total tubule length was doubled, compared with values for the quiescent months. Variable testicular development occurred during transitional intervals preceding and following peak testicular activity. From February to Aril thickening of the seminiferous epithelium and appearance of spermatozoa in the caput epididymidis signalled re-establishment of sperm production. In August shedding of germinal cells from the epithelium heralded impending failure of spermatogenesis. Evidence of an annual testicular cycle contradicted the prevalent belief that equatorial hyrax breed all year and suggested that the testicular cycle is a conservative element of hyracoid reproductive strategy.     

Environmental correlate of the testicular events in a major carp Catla catla in an annual reproductive cycle

Current communication describes annual testicular events in free-living Indian major carp Catla catla and their probable environmental synchronizer(s). The study was initiated with month-wise evaluation of gametogenic and steroidogenic status of the testis, and thus dividing the annual testicular cycle into the preparatory spawning (November to March), the pre-spawning (April to June), the spawning (July to August) and the post-spawning (September to October) phases. An exhaustive statistical analysis of the data on the studied variables of testicular functions and various components of the environment indicated seasonal fluctuations of photoperiod as the major environmental factor associated with the seasonal reproductive activity of this carp. Ambient temperature appeared as a dependent variable of photoperiod, and thereby, may have substantial influences on the development of testis in Catla catla. Rainfall, on the other hand, showed significant correlation only with the peak reproductive activity, i.e. the act of spawning. Collectively, it appears logical to surmise that photo-thermal conditions may act as proximate and rainfall may play a role of ultimate environmental factor in the regulation of annual testicular events in Indian major carp Catla catla.     

Mating-associated peak in plasma testosterone concentration in wild male grey-headed flying foxes (Pteropus poliocephalus)

Plasma testosterone (T) concentrations, measured in wild bats of P. poliocephalus in Queensland in 1983-87, showed a peak during the mating season in March. Plasma androstenedione (A) concentrations changed less dramatically with season. Mean testicular concentration and total content of T and A was substantially greater in March than in regressed testes in July-October. Paired adrenal glands were heavier during February to April than during September to November. In the same wild population, throughout a single breeding season (1987), plasma T concentrations were significantly higher in mid-March than 3 weeks previously or 3 weeks later. Testicular T content rose as the breeding season progressed, being greatest during March, coinciding with the large rise in plasma T concentrations. Testicular T concentration and content were correlated significantly with plasma T concentrations. Adrenal glands contained T, but the absolute concentrations were much lower than in the testis. No significant changes in plasma, testicular or adrenal A concentrations were found as the breeding season progressed. The large increase in plasma T during the mating season appears to be due to increased testicular production.     

Seasonal changes in spermatogenesis in the blue fox (Alopex lagopus), quantified by DNA flow cytometry and measurement of soluble Mn2+ -dependent adenylate cyclase activity

The testes of the blue fox (Alopex lagopus) showed marked seasonal variations in size. Testicular weight and volume increased rapidly during January and February to reach maximal values by the beginning of the breeding season (approximately 15 March). During May and June the weights and volumes of the testes declined gradually to the quiescent state which lasted from July until October. Quantitation by DNA flow cytometry of the seasonal changes in the relative numbers of haploid (1C), diploid (2C) and tetraploid (4C) cell numbers in the testis showed that the increase in testis size from December to February was associated with a rapid expansion of the haploid cell compartment as spermatogenesis resumed. In addition, an increase in number of more mature cell types within the haploid cell population was observed over a 2-month period before the breeding season. The decline in testicular size from the middle of April until October was associated with a reduction in both the absolute and relative sizes of the haploid and tetraploid cell populations and a concomitant increase in the relative numbers of diploid cells. Measurements of the activity of the soluble Mn2+ -dependent adenylate cyclase revealed seasonal variations that closely paralleled those of the haploid cell population, indicating that, as in other species, the enzyme may be associated with maturing germ cells.     

Further observations and considerations on the biological and reproductive cycle of the roe buck (Capreolus capreolus L.)

The authors have studied the seasonal microanatomical modifications of the ovary of the roe deer and testis of the roe buck. The ovary during the month of September presents primary, secondary and mature vesicular follicles. During the month of October there is the first presence of a corpus luteum. The corpora lutea during the months of November and December increase in size and blood supply; the cells are filled with acidophil granules. The corpora lutea persist also in January and February when embryos are implanted in the uterus. This result agrees with Short's and -Hay's but differs from that of Stieve. Testes show the first spermatogenic activity during the month of April. The spermatogenesis is completed in June, persists and reaches the top during July and August, when the Leydig cells acquire a strong acidophil cytoplasm. Indeed the spermatogenesis decreases during September and October; then, during winter months the testes are devoid of all signs of spermatogenic activity, the tubules being lined only be Sertoli cells and spermatogonia.     

Environmental regulation of the reproductive system in a flexibly breeding Sonoran Desert bird, the Rufous-winged Sparrow, Aimophila carpalis

We investigated reproductive regulation in male Rufous-winged Sparrows, Aimophila carpalis, a Sonoran Desert passerine that breeds after irregular summer rains. Field and captive data demonstrate that increased photoperiod stimulates testicular development in March and maintains it until early September. Free-living birds caught in July and placed on captive long days (16L: 8D) maintained developed testes for up to 7 months, and free-living birds caught in September, during testicular regression, redeveloped testes when placed on captive long days, indicating that these birds were still photosensitive. Captive birds on long days maintained testicular development when exposed to temperatures mimicking those occurring during regression in free-living birds. In free-living birds, testicular development was observed during spring and summer, but unless this was associated with rainfall, breeding (indicated by juveniles) did not occur. Large increases in plasma luteinizing hormone (LH) in free-living males were correlated with heavy rainfall in July/August, when the birds bred, and in November, when they did not breed. In captive birds, plasma LH concentrations were unresponsive to photoperiodic changes, but may have responded to social cues. Plasma prolactin concentrations were directly correlated with photoperiod in free-living birds, but an effect of photoperiod on prolactin secretion was not seen in captive birds. It is concluded that male Rufous-winged Sparrows use long photoperiods to stimulate and maintain testicular development, but exposure to long photoperiods does not terminate breeding by inducing absolute photorefractoriness. The specific timing of reproductive behaviors is apparently determined by elevated plasma LH coinciding with long day stimulated gonad development.     

卵胎生硬骨鱼褐菖(鱼良)鲉精巢的周期发育

研究了卵胎生硬骨鱼褐菖(鱼良)鲉(Sebastiscus marmoratus)的精巢结构和生殖周期.褐菖(鱼良)鲉精巢属于小叶型.每年8～9月,精巢处于精原细胞增殖期.初级精原细胞分裂增殖,产生次级精原细胞.后者和支持细胞组成精小囊.10月～翌年1月进入精子发生期.精小囊中的生殖细胞进一步发育,逐渐形成精子.2～7月是精子退化吸收期,精巢中仅有初级精原细胞和残余的精子.在生殖季节,精子经由输出管和输精管运至尿殖突,通过体内受精方式送入雌鱼生殖道.     

Melatonin in the Regulation of Annual Testicular Events in Carp Catla catla: Evidence from the Studies on the Effects of Exogenous Melatonin,Continuous Light,and Continuous Darkness

The physiological significance of melatonin in the regulation of annual testicular events in a major carp Catla catla was evaluated through studies on the effects of graded dose (25, 50, or 100 µg/100 g body wt.) of melatonin exogenously administered for different durations (1, 15, or 30 days) and manipulation of the endogenous melatonin system by exposing the fish to constant darkness (DD) or constant light (LL) for 30 days. An identical experimental schedule was followed during the preparatory (February–March), pre‐spawning (April–May), spawning (July–August), and post‐spawning (September–October) phases of the annual cycle. Irrespective of the reproductive status of the carp, LL suppressed while DD increased the mid‐day and mid‐night values of melatonin compared to respective controls. Influences of exogenous melatonin varied in relation to the dose and duration of treatment and the reproductive status of the carp. However, testicular response to exogenous melatonin (at 100 µg, for 30 days) and DD in each reproductive phase was almost identical. Notably, precocious testicular maturation occurred in both DD and melatonin‐injected fish during the preparatory phase and in LL carps during the pre‐spawning phase. In contrast, testicular functions in both the melatonin‐treated and DD fish were inhibited during the pre–spawning and spawning phases, while the testes did not respond to any treatment during the post‐spawning phase. In conclusion, this study provided the first experimental evidence that melatonin plays a significant role in the regulation of annual testicular events in a sub‐tropical surface‐dwelling carp Catla catla, but the influence of this pineal hormone on the seasonal activity of testis varies in relation to the reproductive status of the concerned fish.     

Seasonal changes in epididymis and the effects of FSH and testosterone in the spiny-tailed lizard,Uromastix hardwicki

Seasonal changes in epididymal weight and histology were studied in relation to testicular function in the adult spiny-tailed lizard, Uromastix hardwicki, over a period of 1 year. The eipdidymal weights, tubular diameter, and epithelial height increased in March, reaching a peak in April. This peak coincided with sperm maturation, elevated plasma testosterone levels, and release of sperm into the epididymis. The epididymal weights decreased in May following a sudden regression of the testis early in the month. The epididymal weights decreased further during June and remained low until February. The diameter of the duct and the height of the epithelial cells also decreased in May and the epididymal epithelium maintained a low histological profile from June to February. The fall testicular recrudescence was not accompanied by a change either in the weight or the histological structure of the epididymis. Administration of oFSH (0.1 mg) daily for 7 days during the sexually quiescent period induced a significant increase in the weight of the epididymis and epithelial height of the duct. Administration of testosterone alone, (2.0 mg) daily for the same period and under identical conditions, did not induce a change in the weight of epididymis or its histology. A possible permissive role of gonadotrophin in the hormonal regulation of the lizard epididymis has been suggested.     

Annual changes in the testicular activity of the river sculpin, Cottus hangiongensis Mori, with emphasis on the occurrence of aberrant spermatids during spermatogenesis

Annual changes in the spermatogenetic activity of the testis were studied histologically in the river sculpin. Coitus hangiongensis , sampled monthly from a river in southern Hokkaido, Japan. A pair of sperm reservoirs, consisting of many anastomosing lacunae, was present along the dorsomedian edge of the paired testes, and functioned also as a sperm-transporting system instead of the typical sperm duct. Spermatogenesis occurred actively in August, yielding an increasing number of mature spermatozoa in October. This process advanced, but slowly during the succeeding winter months, until March. The testis became functionally mature during the spawning period in April and May. In July, small numbers of spermatocytes were found to have appeared already, which indicated a relatively short period of post-spawning testicular regression. In November, germinal cysts containing aberrant binuclear spermatids began to appear within the seminal lobules. The paired nuclei of aberrant spermatids gradually enlarged, and the cells were released into the lumina of the seminal lobules simultaneously with the release of mature spermatozoa from the germinal cysts. During the functional maturity stage, lumina of seminal lobules which had expelled mature spermatozoa to sperm reservoirs became filled with these abnormal bodies. Discussion includes the occurrence of aberrant spermatids which resulted in the formation of 'spermatid masses' as has been described in other cottids.     

Control of lake trout reproduction: role of lipids

Somatic lipid density of lake trout decreased during summer and increased after spawning in October, peaking April, prior to ovarian recrudescence. Non-lipid density increased from June or July until August and remained relatively constant until March. All lake trout ( n =30) reared on high ( n =10, 0·4%), medium ( n =10, 0·15%) and low ( n =10, 0·05%) rations from March to September spawned. In a second experiment for which the treatments commenced in November, the proportion of females maturing in the following October (high=88%, medium=69%, low=28%) was positively related to ration levels.