6 research outputs found
Sperm morphological abnormalities in autosomal dominant polycystic kidney disease are associated with the Hippo signaling pathway via PC1
BackgroundAutosomal dominant polycystic kidney disease (ADPKD) is a hereditary kidney disorder mostly caused by mutations in PKD1 or PKD2 genes. Here, we report thirteen ADPKD males with infertility and investigated the sperm morphological defects associated with PC1 disruption.MethodsTargeted next-generation sequencing was performed to detect PKD1 variants in patients. Sperm morphology was observed by immunostaining and transmission electron microscopy, and the sperm motility was assessed using the computer-assisted sperm analysis system. The Hippo signaling pathway was analyzed with by quantitative reverse transcription polymerase chain reaction (qPCR) and western blotting in vitro.ResultsThe ADPKD patients were infertile and their sperm tails showed morphological abnormalities, including coiled flagella, absent central microtubules, and irregular peripheral doublets. In addition, the length of sperm flagella was shorter in patients than in controls of in in. In vitro, ciliogenesis was impaired in Pkd1-depleted mouse kidney tubule cells. The absence of PC1 resulted in a reduction of MST1 and LATS1, leading to nuclear accumulation of YAP/TAZ and consequently increased transcription of Aurka. which might promote HDAC6-mediated ciliary disassembly.ConclusionOur results suggest the dysregulated Hippo signaling significantly contributes to ciliary abnormalities in and may be associated with flagellar defects in spermatozoa from ADPKD patients
Autosomal Dominant Polycystic Kidney Disease: Extrarenal Involvement
: Autosomal dominant polycystic kidney disease (ADPKD) is the most common hereditary kidney disorder, but kidneys are not the only organs involved in this systemic disorder. Individuals with the condition may display additional manifestations beyond the renal system, involving the liver, pancreas, and brain in the context of cystic manifestations, while involving the vascular system, gastrointestinal tract, bones, and cardiac valves in the context of non-cystic manifestations. Despite kidney involvement remaining the main feature of the disease, thanks to longer survival, early diagnosis, and better management of kidney-related problems, a new wave of complications must be faced by clinicians who treated patients with ADPKD. Involvement of the liver represents the most prevalent extrarenal manifestation and has growing importance in the symptom burden and quality of life. Vascular abnormalities are a key factor for patients' life expectancy and there is still debate whether to screen or not to screen all patients. Arterial hypertension is often the earliest onset symptom among ADPKD patients, leading to frequent cardiovascular complications. Although cardiac valvular abnormalities are a frequent complication, they rarely lead to relevant problems in the clinical history of polycystic patients. One of the newest relevant aspects concerns bone disorders that can exert a considerable influence on the clinical course of these patients. This review aims to provide the "state of the art" among the extrarenal manifestation of ADPKD
Enhanced fertility estimation and novel ion channels in boar sperm
Cryopreserved boar sperm have many potential advantages when used for artificial insemination (AI), including increased time for pathogen testing and ease of semen transport. However, frozen boar sperm are seldom used for AI due to decreased pregnancy rates and litter sizes as a result of cryo-induced damage to sperm. Estimating the fertility of samples prior to AI would allow disposal of semen with poor predicted fertility. Unfortunately, conventional laboratory analyses of semen do not provide accurate estimates of fertility. Assessing sperm traits that more accurately predict fertility would improve commercial use of frozen boar sperm. Our objective was to identify traits of cryopreserved sperm that are related to boar fertility by including novel sperm assays with traditional laboratory analyses that may collectively provide a more accurate model for estimating fertility. Semen from 14 boars of several breeds was cooled to 15°C for overnight shipping prior to freezing. Semen was thawed and motility was estimated and confirmed using Computer Automated Sperm Assessment (CASA). Sperm viability and acrosome integrity were measured at 0, 30 and 60 min post-thaw. In addition to traditional analyses, each sperm sample was tested by IVF in two to three independent replicates and fertilization, cleavage and blastocyst development were recorded. As an assessment of sperm reservoir formation, a sperm-oviduct binding assay was used to compare the number of sperm bound to epithelial aggregates harvested from gilt isthmus. Additionally, a competitive zona binding assay using two distinct fluorophores for sperm identification was employed to measure the number of sperm from each boar bound to the zona. Frozen sperm from the same ejaculates subjected to laboratory analyses were used to determine boar fertility. Fertility was measured by AI of mature gilts using 4.0 x 106 sperm from one boar at 24 h and a second boar at 36 h after the onset of estrus and calculated as the percentage of the litter sired by each boar. AI order was reversed in consecutive replicates so that order of insemination was evenly distributed among boar comparisons. Reproductive tracts were harvested at ~ 32 d after AI and the number of fetuses were recorded and sampled for paternity identification using microsatellite markers. The least-squared means (LSM) of each laboratory evaluation were modeled by boar using multiple regression analyses to test their collective values in predicting fertility. The model generated was highly predictive of fertility (P < 0.001, r2 = 0.87) and included 5 traits; acrosome compromised sperm (0 and 30 min), percent live sperm (0 min), percent total motility (30 min) and the number of zona bound sperm. An additional model in which fertility was assessed by the number of piglets sired by boar also predicted fertility (P < 0.05, r2 = 0.57) and shared many of the same traits including percent live sperm, motility and the number of sperm bound to zona. These models indicate that the fertility of cryopreserved boar sperm can be predicted using both traditional and novel laboratory assays that consider multiple functions of sperm. This model provides a method to cull low fertility cryopreserved boar semen more accurately, thus enhancing the potential adoption of semen cryopreservation by the swine industry.
Both models that successfully estimated boar fertility included sperm traits that evaluated binding ability to the zona pellucida. Further, binding to the oviduct epithelium was related to fertility. Impaired functions of sperm membrane proteins that are critical for normal sperm interactions within the female reproductive tract are not always measured during routine semen analyses and therefore likely contribute to idiopathic subfertility. Sperm must undergo important capacitation-like changes for normal fertilization such as a change in motility parameters and an increase in intracellular calcium. Transient receptor potential channel-2 (TRPP2) is a membrane protein with regulatory roles in ion homeostasis. TRPP2 is important for sperm motility, storage and male fertility in lower organisms (Drosophila melanogaster), but the protein has not been described in mammalian sperm. My objective was to identify, localize and begin to determine the functions of TRPP2 in porcine sperm. TRPP2 was detected in both capacitated and non-capacitated boar sperm and was localized to the head and principle piece. Antisera to TRPP2 inhibited sperm movement and increased tail cross-beat frequency. Sperm incubated with TRPP2 antibody had lower levels of free intracellular calcium and did not experience a rise in calcium over time when maintained in capacitating conditions. We conclude that porcine TRPP2 is a previously unreported mammalian sperm ion channel that regulates capacitation-like changes in boar sperm
Removal of antagonistic spindle forces can rescue metaphase spindle length and reduce chromosome segregation defects
Regular Abstracts - Tuesday Poster Presentations: no. 1925Metaphase describes a phase of mitosis where chromosomes are attached and oriented on the bipolar spindle for subsequent segregation at anaphase. In diverse cell types, the metaphase spindle is maintained at a relatively constant length. Metaphase spindle length is proposed to be regulated by a balance of pushing and pulling forces generated by distinct sets of spindle microtubules and their interactions with motors and microtubule-associated proteins (MAPs). Spindle length appears important for chromosome segregation fidelity, as cells with shorter or longer than normal metaphase spindles, generated through deletion or inhibition of individual mitotic motors or MAPs, showed chromosome segregation defects. To test the force balance model of spindle length control and its effect on chromosome segregation, we applied fast microfluidic temperature-control with live-cell imaging to monitor the effect of switching off different combinations of antagonistic forces in the fission yeast metaphase spindle. We show that spindle midzone proteins kinesin-5 cut7p and microtubule bundler ase1p contribute to outward pushing forces, and spindle kinetochore proteins kinesin-8 klp5/6p and dam1p contribute to inward pulling forces. Removing these proteins individually led to aberrant metaphase spindle length and chromosome segregation defects. Removing these proteins in antagonistic combination rescued the defective spindle length and, in some combinations, also partially rescued chromosome segregation defects. Our results stress the importance of proper chromosome-to-microtubule attachment over spindle length regulation for proper chromosome segregation.postprin
Psr1p interacts with SUN/sad1p and EB1/mal3p to establish the bipolar spindle
Regular Abstracts - Sunday Poster Presentations: no. 382During mitosis, interpolar microtubules from two spindle pole bodies (SPBs) interdigitate to create an antiparallel microtubule array for accommodating numerous regulatory proteins. Among these proteins, the kinesin-5 cut7p/Eg5 is the key player responsible for sliding apart antiparallel microtubules and thus helps in establishing the bipolar spindle. At the onset of mitosis, two SPBs are adjacent to one another with most microtubules running nearly parallel toward the nuclear envelope, creating an unfavorable microtubule configuration for the kinesin-5 kinesins. Therefore, how the cell organizes the antiparallel microtubule array in the first place at mitotic onset remains enigmatic. Here, we show that a novel protein psrp1p localizes to the SPB and plays a key role in organizing the antiparallel microtubule array. The absence of psr1+ leads to a transient monopolar spindle and massive chromosome loss. Further functional characterization demonstrates that psr1p is recruited to the SPB through interaction with the conserved SUN protein sad1p and that psr1p physically interacts with the conserved microtubule plus tip protein mal3p/EB1. These results suggest a model that psr1p serves as a linking protein between sad1p/SUN and mal3p/EB1 to allow microtubule plus ends to be coupled to the SPBs for organization of an antiparallel microtubule array. Thus, we conclude that psr1p is involved in organizing the antiparallel microtubule array in the first place at mitosis onset by interaction with SUN/sad1p and EB1/mal3p, thereby establishing the bipolar spindle.postprin
Мужское бесплодие: этиология, патогенез, классификация, диагностика и методы лечения
В монографии раскрыты вопросы репродуктивной физиологии человека,
гормональной регуляции сперматогенеза. Изложена информация об этиопатогенезе и
механизмах развития мужского бесплодия. Раскрыты вопросы диагностики мужского
бесплодия, а также все виды бесплодия (эндокринное, секреторное, обструктивное,
иммунологическое, гормональное, тестикулярная недостаточность, генетические нарушения,
варикоцеле, крипторхизм, идиопатическое бесплодие, бесплодие при заболеваниях,
передающихся половым путем). Освещены вопросы вклада мужчин в контрацепцию.
Изложены общие принципы лечения бесплодия у мужчин, лечения мужского фактора
бесплодия супружеской пары, а также направления профилактики мужского бесплодия.
Монография предназначена для андрологов, урологов, эндокринологов,
сексопатологов, врачей общей практики и врачей других специальностей, а также будет
полезной для клинических ординаторов, врачей-интернов и студентов медицинских вузов