Role of follitropin receptor signaling in nuclear protein transitions and chromatin condensation during spermatogenesis

Follitropin receptor (FSHR) in testicular Sertoli cells mediates signaling by pituitary follitropin (FSH) promoting intercellular communication with germ cells for normal spermatogenesis. Using receptor knockout mice we examined changes in sperm nucleoproteins and chromatin architecture. The expressions of transition proteins 1/2 (TP1/2) and protamine-2 (PRM-2) were greatly diminished at 21 days, but returned to normal at 35 days and 3 months after birth. However, protein components in chromatin were quite different. Western blots detected a reduction in PRM1/2 and prolonged retention of mono-ubiquitinated histone 2A (uH2A) in the epididymal sperm from adult mutants. Two forms of mono- and poly-uH2A were present in sonication-resistant testicular spermatids in normal mice, whereas only an elevated mono-uH2A was detectable in mutants. Decrease in PRM1/2 and retention of mono-uH2A was coincident with reduction in TP1/2 in premature spermatids. Thus lack of FSHR signaling impairs expression of TP1/2 and PRM-2 at an early stage of post-natal development causing delayed spermatogenesis. In the adult, absence of FSHR signaling prolongs retention of mono-uH2A, leading to impair transition of basic nucleoproteins and chromatin remodeling during mouse spermatogenesis.     

Endocrine alterations and signaling changes associated with declining ovarian function and advanced biological aging in follicle-stimulating hormone receptor haploinsufficient mice

Reproductive aging in female mammals is characterized by a progressive decline in fertility due to loss of follicles and reduced ovarian steroidogenesis. In this study we examined some of the endocrine and signaling parameters that might contribute to a decrease in ovulation and reproductive performance of mice with haploinsufficiency of the FSH receptor (FSH-R). For this purpose we compared ovarian changes and hormone levels in FSH-R heterozygous (+/-) and wild-type mice of different ages (3, 7, and 12 mo). Hormone-induced ovulations in immature and 3-mo-old +/- mice were consistently lower. The number of corpora lutea (CL) were lower at 3 and 7 mo, and none were present in 1-yr-old +/- females. The plasma steroid and gonadotropin levels exhibited changes associated with typical ovarian aging. Plasma FSH and LH levels were higher in 7-mo-old +/- mice, but FSH levels continued to rise in both genotypes by 1 yr. Serum estradiol and progesterone were lower in +/- mice at all ages, and testosterone was several-fold higher in 7-mo-old and 1-yr-old +/- mice. Inhibin alpha (Western blot) appeared to be lower in +/- ovaries at all ages. FSH-R (FSH* binding) declined steadily from 3 mo and reaching the lowest point at 1 yr. LH receptor (LH* binding) was high in the 1-yr-old ovary, and expression was localized in the stroma and interstitial cells. Our findings demonstrate that haploinsufficiency of the FSH-R gene could cause premature exhaustion of the gonadal reserves previously noted in these mice. This is accompanied by age-related changes in the hypothalamic-pituitary axis. As these features in our FSH-R +/- mice resemble reproductive failure occurring in middle-age women, further studies in this model might provide useful insights into the mechanisms underlying ovarian aging.     

Effect of Piper longum Linn, Zingiber officianalis Linn and Ferula species on gastric ulceration and secretion in rats

Use of Dipaniya Mahakasaya, a group consisting of 10 herbal drugs, has been suggested in Charaka Samhita to improve digestion. Out of these 10 plants, three, viz. P. longum (water decoction), Z. officianalis (water decoction) and Ferula species (colloidal solution) were studied for their antiulcer and mechanism of antiulcer effects in rats. All the drugs in the dose of 50 mg/kg, p.o., 60 min prior to experiment, showed significant protection against gastric ulcers induced by 2 hr cold restraint stress, aspirin (200 mg/kg, 4 hr) and 4 hr pylorus ligation. The antiulcerogenic effect seemed to be due to the augmentation of mucin secretion and decreased cell shedding rather than offensive acid and pepsin secretion which however, were found to be increased by them.     

Qualitative and quantitative decline in spermatogenesis of the follicle-stimulating hormone receptor knockout (FORKO) mouse

Sertoli cells express functional receptors for FSH, one of the two pituitary hormones that regulate spermatogenesis in mammals. We recently produced genetic mutant (FORKO) mice that lack FSH receptor, in order to examine the effects on testicular function and fertility. Mutant males exhibited weight loss of testis, epididymis, and seminal vesicle as well as low levels of testosterone. Except for reduced seminiferous tubular diameter, no gross changes were apparent upon histological examination. Analysis of testicular germ cells by flow cytometry revealed a significant increase in the percentage of 2C cells (spermatogonia and non-germ cells) and a significant decrease in the percentage of HC cells (elongated spermatids) of FORKO males. The absolute number of homogenization-resistant elongated spermatids was also significantly reduced in the mutant males. A 2-fold increase in c-kit-positive 2C cells was recorded in the mutant males. Elongated spermatids of FORKO males showed a dramatic increase in propidium iodide binding suggesting reduced nuclear compaction. The increase in size of the sperm head in mutants, as well as susceptibility to dithiothreitol-induced decondensation, suggests the inadequate condensation of sperm chromatin. Sperm chromatin structure assay, a technique that reflects DNA stability, revealed that sperm from FORKO males are susceptible to acid denaturation, indicating the poor quality of sperm. These data allow us to conclude that genetic disruption of FSH receptor signaling in the rodent induces major changes that might contribute to reduced fertility.     

The emergence of okra enation leaf curl virus—an important begomovirus,infecting okra in several states across India

Okra enation leaf curl virus (OELCuV), a begomovirus, is an emerging serious constraint for okra production in India. OELCuV was earlier reported in Haryana, Gujarat and Karnataka. In the current study, a survey was conducted on okra crop in the other predominant okra growing regions of India and identified for the first time, a widespread symptomology of OELCuV. The disease incidence was recorded between 5 and 74% in all the surveyed regions. The diseased samples collected from all the locations were amplified with coat protein specific new primer in PCR. The amplicons were sequenced and deposited to NCBI Gene Bank. The finding could be highly useful in okra breeding programs against OELCuV.     

Regeneration of plants from seed-derived callus of Hybanthus enneaspermus L. Muell., a rare ethnobotanical herb

A procedure was developed for plant regeneration of Hybanthus enneaspermus, a rare ethnobotanical herb from the Deccan peninsula in India, through seed-derived callus. Seeds demonstrated a high induction frequency (69.4±2.8%) and a high yield (364.4±2.5 mg) of light-yellow friable callus on Murashige and Skoog's (MS) medium containing 2.6 μm NAA and 2.2 μm BA within 4 weeks of incubation. After 1 year of subculture, yellow friable and light-green compact calli types were established from initial light-yellow friable callus. Shoot differentiation was achieved from light-green compact callus, but not from yellow friable callus. Shoot differentiation resulted when light-green compact callus was transferred to MS medium supplemented with 8.8 μm BA and 2.6 μm NAA; the highest percentage of calli forming shoots (66.6±4.8%) and the highest number of shoots (8.9±0.3) were achieved in this medium. Differentiated shoot buds elongated to 4–5 cm within 4 weeks. The addition of casein hydrolysate (500 mg/l) and more potassium phosphate (1.86 mm) to the culture medium enhanced shoot differentiation. Rooting was achieved on the shoots using half-strength MS medium containing 4.8 μm IBA. About 70% of the plants were established in pots containing pure garden soil after 2 weeks of hardening. The regenerated plants were morphologically uniform and exhibited normal seed set. Received: 23 July 1998 / Revision received: 18 November 1998 / Accepted: 26 November 1998     

Induction of Oxidative Stress and Antioxidant Activity by Hydrogen Peroxide Treatment in Tolerant and Susceptible Wheat Genotypes

We induced an oxidative stress by means of exogenous hydrogen peroxide in two wheat genotypes, C 306 (tolerant to water stress) and Hira (susceptible to water stress), and investigated oxidative injury and changes in antioxidant enzymes activity. H₂O₂ treatment caused chlorophyll degradation, lipid peroxidation, decreased membrane stability and activity of nitrate reductase. Hydrogen peroxide increased the activity of antioxidant enzymes, glutathione reductase and catalase. These effects increased with increasing H₂O₂ concentrations. However, no change was observed in the activity of superoxide dismutase and proline accumulation.     

Increased Antioxidant Activity under Elevated Temperatures: A Mechanism of Heat Stress Tolerance in Wheat Genotypes

An experiment was conducted with three wheat (Triticum aestivum L.) genotypes C 306, HD 2285 and HD 2329 (differently susceptible to water and temperature stress) to study the extent of oxidative injury and activities of antioxidant enzymes in relation to heat stress induced by manipulating dates of sowing. Increase in temperature by late sowing significantly decreased leaf relative water content (RWC), ascorbic acid content, and increased H₂O₂ content and lipid peroxidation in all the genotypes at 8 and 23 d after anthesis. Temperature tolerant genotypes C 306, closely followed by HD 2285 were superior to HD 2329 in maintaining high RWC, ascorbic acid content, and lower H₂O₂ content and lipid peroxidation (malondialdehyde content) under high temperature (late sowing) at the two stages. Activities of superoxide dismutase and catalase were highest in HD 2285 followed by C 306 and minimum in HD 2329 while ascorbate peroxidase activity was highest in C 306.