The microtubule organizing centers(MTOCs)of human and mouse oocytes are essential for meiotic spindle assembly and for ensuring precise chromosome segregations.Previous studies mainly focus on investigating MTOCs chan...The microtubule organizing centers(MTOCs)of human and mouse oocytes are essential for meiotic spindle assembly and for ensuring precise chromosome segregations.Previous studies mainly focus on investigating MTOCs changes in metaphase I oocyte.However,the detailed dynamic changes and underlying mechanisms of the MTOCs in germinal vesicle(GV)oocytes—a stage that early events of MTOC maturation happened—remain unclear.Here we explored the dynamics of MTOCs maturation in human and mouse GV oocytes and found that MTOCs maturation is a largely conserved process,consisting of two tightly coupled processes referred to as MTOCs activation and migration.We found that cytoskeleton associated protein 5(CKAP5)and transforming acidic coiled-coil containing protein 3(TACC3)play key roles in MTOCs maturation in oocytes.The activation of the MTOCs is a prerequisite for migration initiation,and the migration of the MTOCs is facilitated by dynein/dynactin in oocytes.The disruption of MTOC maturation resulted in spindle assembly failure.Importantly,impaired MTOCs maturation is associated with the physiological aging of oocytes.This study provides a comprehensive understanding of MTOCs dynamics in human and mouse oocytes.展开更多
Dear Editor,Previously,the Mendelian phe no types in huma n oocyte maturation arrest,fertilization failure and early embryonic arrest,are largely underestimated.In recent years,"missing"Men delian phe no typ...Dear Editor,Previously,the Mendelian phe no types in huma n oocyte maturation arrest,fertilization failure and early embryonic arrest,are largely underestimated.In recent years,"missing"Men delian phe no types and genes in these processes are beginning to be uncovered by us and others(Huang et al.,2014;Alazami et al..2015;Feng et al.,2016;Xu et al.,2016;Chen et al.,2017;Sang et al.,2019).However,the genetic basis for majority of patients resulting from abnormalities in these phe no types remains to be elucidated.展开更多
Teratozoospermia is usually associated with defective spermiogenesis and is a disorder with considerable genetic heterogeneity. Although previous studies have identified several teratozoospermia-associated genes, the ...Teratozoospermia is usually associated with defective spermiogenesis and is a disorder with considerable genetic heterogeneity. Although previous studies have identified several teratozoospermia-associated genes, the etiology remains unknown for a majority of affected men. Here, we identified a homozygous missense mutation and a compound heterozygous mutation of CCIN in patients suffering from teratozoospermia. CCIN encodes the cytoskeletal protein Calicin that is involved in the formation and maintenance of the highly regular organization of the calyx of mammalian spermatozoa, and has been proposed to play a role in sperm head structure remodeling during the process of spermiogenesis. Our morphological and ultrastructural analyses of the spermatozoa obtained from all three men harboring deleterious CCIN mutants reveal severe head malformation. Further immunofluorescence assays unveil markedly reduced levels of Calicin in spermatozoa. These patient phenotypes are successfully recapitulated in mouse models expressing the disease-associated variants, confirming the role of Calicin in male fertility.Notably, all mutant spermatozoa from mice and human patients fail to adhere to the zona mass, which likely is the major mechanistic reason for CCIN-mutant sperm-derived infertility. Finally, the use of intracytoplasmic sperm injections(ICSI) successfully makes mutated mice and two couples with CCIN variants have healthy offspring. Taken together, our findings identify the role of Calicin in sperm head shaping and male fertility, providing important guidance for genetic counseling and assisted reproduction treatments.展开更多
Clustered protocadherins(Pcdhs)are a large family of cadherin-like cell adhesion proteins that are central for neurite selfavoidance and neuronal connectivity in the brain.Their downstream nonreceptor tyrosine kinase ...Clustered protocadherins(Pcdhs)are a large family of cadherin-like cell adhesion proteins that are central for neurite selfavoidance and neuronal connectivity in the brain.Their downstream nonreceptor tyrosine kinase Pyk2(proline-rich tyrosine kinase 2,also known as Ptk2b,Cakb,Raftk,Fak2,and Cadtk)is predominantly expressed in the hippocampus.We constructed Pyk2-null mouse lines and found that these mutant mice showed enhancement in contextual fear memory,without significant change in auditory-cued and spatial-referenced learning and memory.In addition,by preparing Y402F mutant mice,we observed that Pyk2 suppressed contextual fear memory in an autophosphorylation-independent manner.Moreover,using high-throughput RNA sequencing,we found that immediate early genes,such as Npas4,cFos,Zif268/Egr1,Arc,and Nr4a1,were enhanced in Pyk2-null mice.We further showed that Pyk2 disruption affected pyramidal neuronal complexity and spine dynamics.Thus,we demonstrated that Pyk2 is a novel fear memory suppressor molecule and Pyk2-null mice provide a model for understanding fearrelated disorders.These findings have interesting implications regarding dysregulation of the Pcdh–Pyk2 axis in neuropsychiatric disorders.展开更多
基金supported by the National Natural Science Foundation of China(82325021,82288102,32130029,82422033,82271685,82171643,and 82171685)the Shanghai Rising-Star Program(23QA1401800)+3 种基金the New Cornerstone Science Foundation through the XPLORER PRIZE,the Fund of Fudan University and Cao’ejiang Basic Research(24FCB01)the SANS Exploration Scholarthe Fund for Excellent Young Scholars of Shanghai Ninth People’s Hospital,Shanghai Jiao Tong University School of Medicine(JYYQ004)the Shanghai JIAI Genetics and IVF Institute Found(JA-2025033).
文摘The microtubule organizing centers(MTOCs)of human and mouse oocytes are essential for meiotic spindle assembly and for ensuring precise chromosome segregations.Previous studies mainly focus on investigating MTOCs changes in metaphase I oocyte.However,the detailed dynamic changes and underlying mechanisms of the MTOCs in germinal vesicle(GV)oocytes—a stage that early events of MTOC maturation happened—remain unclear.Here we explored the dynamics of MTOCs maturation in human and mouse GV oocytes and found that MTOCs maturation is a largely conserved process,consisting of two tightly coupled processes referred to as MTOCs activation and migration.We found that cytoskeleton associated protein 5(CKAP5)and transforming acidic coiled-coil containing protein 3(TACC3)play key roles in MTOCs maturation in oocytes.The activation of the MTOCs is a prerequisite for migration initiation,and the migration of the MTOCs is facilitated by dynein/dynactin in oocytes.The disruption of MTOC maturation resulted in spindle assembly failure.Importantly,impaired MTOCs maturation is associated with the physiological aging of oocytes.This study provides a comprehensive understanding of MTOCs dynamics in human and mouse oocytes.
文摘Dear Editor,Previously,the Mendelian phe no types in huma n oocyte maturation arrest,fertilization failure and early embryonic arrest,are largely underestimated.In recent years,"missing"Men delian phe no types and genes in these processes are beginning to be uncovered by us and others(Huang et al.,2014;Alazami et al..2015;Feng et al.,2016;Xu et al.,2016;Chen et al.,2017;Sang et al.,2019).However,the genetic basis for majority of patients resulting from abnormalities in these phe no types remains to be elucidated.
基金supported by the National Natural Science Foundation of China(31930063,81771533,81901531,31971137,and 81871163)the National Key Research and Development Program of China(2018YFC2000102,2018YFA0107004,and 2018YFC1003000)+2 种基金the Shanghai Municipal Health Commission and Collaborative Innovation Cluster Project(2019CXJQ01)SHIPM-pi Fund(JY201801)SHIPM-mu Fund(JC201802)。
文摘Teratozoospermia is usually associated with defective spermiogenesis and is a disorder with considerable genetic heterogeneity. Although previous studies have identified several teratozoospermia-associated genes, the etiology remains unknown for a majority of affected men. Here, we identified a homozygous missense mutation and a compound heterozygous mutation of CCIN in patients suffering from teratozoospermia. CCIN encodes the cytoskeletal protein Calicin that is involved in the formation and maintenance of the highly regular organization of the calyx of mammalian spermatozoa, and has been proposed to play a role in sperm head structure remodeling during the process of spermiogenesis. Our morphological and ultrastructural analyses of the spermatozoa obtained from all three men harboring deleterious CCIN mutants reveal severe head malformation. Further immunofluorescence assays unveil markedly reduced levels of Calicin in spermatozoa. These patient phenotypes are successfully recapitulated in mouse models expressing the disease-associated variants, confirming the role of Calicin in male fertility.Notably, all mutant spermatozoa from mice and human patients fail to adhere to the zona mass, which likely is the major mechanistic reason for CCIN-mutant sperm-derived infertility. Finally, the use of intracytoplasmic sperm injections(ICSI) successfully makes mutated mice and two couples with CCIN variants have healthy offspring. Taken together, our findings identify the role of Calicin in sperm head shaping and male fertility, providing important guidance for genetic counseling and assisted reproduction treatments.
基金supported by grants from the National Natural Science Foundation of China(31200825 to L.S.and 31630039 to Q.W.)the Ministry of Science and Technology of China(2017YFA0504203 and 2018YFC1004504)+2 种基金the Science and Technology Commission of Shanghai Municipality(19JC1412500 and 21DZ2210200 to Q.W.)Shanghai Jiao Tong University Scientific and Technological Innovation Funds(17JCYB12 to L.S.)Q.W.is a Shanghai Subject Chief Scientist.
文摘Clustered protocadherins(Pcdhs)are a large family of cadherin-like cell adhesion proteins that are central for neurite selfavoidance and neuronal connectivity in the brain.Their downstream nonreceptor tyrosine kinase Pyk2(proline-rich tyrosine kinase 2,also known as Ptk2b,Cakb,Raftk,Fak2,and Cadtk)is predominantly expressed in the hippocampus.We constructed Pyk2-null mouse lines and found that these mutant mice showed enhancement in contextual fear memory,without significant change in auditory-cued and spatial-referenced learning and memory.In addition,by preparing Y402F mutant mice,we observed that Pyk2 suppressed contextual fear memory in an autophosphorylation-independent manner.Moreover,using high-throughput RNA sequencing,we found that immediate early genes,such as Npas4,cFos,Zif268/Egr1,Arc,and Nr4a1,were enhanced in Pyk2-null mice.We further showed that Pyk2 disruption affected pyramidal neuronal complexity and spine dynamics.Thus,we demonstrated that Pyk2 is a novel fear memory suppressor molecule and Pyk2-null mice provide a model for understanding fearrelated disorders.These findings have interesting implications regarding dysregulation of the Pcdh–Pyk2 axis in neuropsychiatric disorders.
