Male Line Research Articles

Carlos II of Spain, ‘The Bewitched’: cursed by aspartylglucosaminuria?

Carlos II of Spain (1661–1700), last of the Spanish Habsburgs, was known as The ‘Bewitched’ due to his multiple medical issues and feeble nature. He suffered from a range of...

Growth Performance of Male Line of Superior Tropical Rabbit

New Zealand White rabbits (NN) are known as broiler rabbits that are well adapted to the Indonesian environment. HyLa rabbits are rabbits imported from China that were selected for high reproduction and growth rates. The increased productivity of NN rabbits was achieved by crossing to HyLa rabbits. This study was aimed to evaluate HyLa and NN rabbits’ productivity and their reciprocal crosses to develop a broiler rabbit adaptive to tropical climate. Forty heads of does and 20 heads of bucks of HyLa rabbit, 30 heads of does, and 6 heads of bucks of New Zealand White rabbits (NN) were used. Reproductive performance of does (litter size at birth, litter size at weaning, mortality, and weekly does body weight) and the kit’s growth performance (weekly body weight from weaning until the age of 20 weeks) was evaluated. Data were analyzed using the SAS program, FactoMineR, and factoextra package in R 4.0.0. Rabbit’s growth data were periodically analyzed by the Gompertz model. At the age of 10 weeks, HyLa had a body weight of 1459.7 ± 373.1 g greater than HyLa x NN (1380.1 ± 389.9 g), NN x HyLa (1225.7 ± 263.4 g), and NN (985.0 ± 234.4 g). Based on the Gompertz model, NN x HyLa had an inflection point at a weight of 1,977.7 at 12 weeks. Furthermore, NN x HyLa had a better litter size than HyLa x NN. NN bucks x HyLa does have the potential to be developed as a superior rabbit adaptive to tropical climate.

Identification and Characterization of the Masculinizing Function of the Helicoverpa armigera Masc Gene

The Masculinizer (Masc) gene has been known to control sex development and dosage compensation in lepidopterans. However, it remains unclear whether its ortholog exists and plays the same roles in distantly related lepidopterans such as Helicoverpa armigera. To address this question, we cloned Masc from H. armigera (HaMasc), which contains all essential functional domains of BmMasc, albeit with less than 30% amino acid sequence identity with BmMasc. Genomic PCR and qPCR analyses showed that HaMasc is a Z chromosome-linked gene since its genomic content in males (ZZ) was two times greater than that in females (ZW). RT-PCR and RT-qPCR analyses revealed that HaMasc expression was sex- and stage-biased, with significantly more transcripts in males and eggs than in females and other stages. Transfection of a mixture of three siRNAs of HaMasc into a male embryonic cell line of H. armigera led to the appearance of female-specific mRNA splicing isoforms of H. armigera doublesex (Hadsx), a downstream target gene of HaMasc in the H. armigera sex determination pathway. The knockdown of HaMasc, starting from the third instar larvae resulted in a shift of Hadsx splicing from male to female isoforms, smaller male pupa and testes, fewer but larger/longer spermatocytes and sperm bundles, delayed pupation and internal fusion of the testes and follicles. These data demonstrate that HaMasc functions as a masculinizing gene in the H. armigera sex-determination cascade.

Isolation and Characterization of APETALA3 Orthologs and Promoters from the Distylous Fagopyrum esculentum.

Common buckwheat (Fagopyrum esculentum) produces distylous flowers with undifferentiated petaloid tepals, which makes it obviously different from flowers of model species. In model species Arabidopsis, APETALA3 (AP3) is expressed in petal and stamen and specifies petal and stamen identities during flower development. Combining with our previous studies, we found that small-scale gene duplication (GD) event and alternative splicing (AS) of common buckwheat AP3 orthologs resulted in FaesAP3_1, FaesAP3_2 and FaesAP3_2a. FaesAP3_2 and FaesAP3_2a were mainly expressed in the stamen of thrum and pin flower. Promoters functional analysis suggested that intense GUS staining was observed in the whole stamen in pFaesAP3_2::GUS transgenic Arabidopsis, while intense GUS staining was observed only in the filament of stamen in pFaesAP3_1::GUS transgenic Arabidopsis. These suggested that FaesAP3_1 and FaesAP3_2 had overlapping functions in specifying stamen filament identity and work together to determine normal stamen development. Additionally, FaesAP3_2 and FaesAP3_2a owned the similar ability to rescue stamen development of Arabidopsis ap3-3 mutant, although AS resulted in a frameshift mutation and consequent omission of the complete PI-derived motif and euAP3 motif of FaesAP3_2a. These suggested that the MIK region of AP3-like proteins was crucial for determining stamen identity, while the function of AP3-like proteins in specifying petal identity was gradually obtained after AP3 Orthologs acquiring a novel C-terminal euAP3 motif during the evolution of core eudicots. Our results also provide a clue to understanding the early evolution of the functional specificity of euAP3-type proteins involving in floral organ development in core eudicots, and also suggested that FaesAP3_2 holds the potential application for biotechnical engineering to develop a sterile male line of F. esculentum.

Selective estrogen receptor modulator, tamoxifen, inhibits Zika virus infection.

Zika virus (ZIKV) is an arbovirus belonging to the flaviviridae family with a risk assessment that has been increasing in recent years and was labeled a global health emergency by the World Health Organization in 2016. There are currently no Food and Drug Administration-approved treatment options available for ZIKV, so expeditious development of treatment options is urgent. To expedite this process, an on-market drug, tamoxifen (TAM), was selected as a promising candidate for repurposing due to its wide range of biological activities and because it has already been shown to possess activity against hepatitis C virus, a flavivirus in a separate genus. Anti-ZIKV activity of TAM was assessed by compound screens using an infectious virus and mechanistic details were gleaned from time of addition and virucidal studies. TAM and an active metabolite, 4-hydroxytamoxifen (TAM-OH), both showed promising antiviral activity (EC50 ≈9 and 5 µM, respectively) in initial compound screening and up to 8-h postinfection, though the virucidal assay indicated that they do not possess any direct virucidal activity. Additionally, TAM was assessed for its activity against ZIKV in the human male germ cell line, SEM-1, due to the sexually transmitted nature of ZIKV owing to its extended survival times in germ cells. Virus titers show diminished replication of ZIKV over 7 days compared to controls. These data indicate that TAM has the potential to be repurposed as an anti-ZIKV therapeutic and warrants further investigation.

From Croatia to Transylvania

The article gives the history of the noble Croatian family of Perušić, following the life and career of its main male representatives across three generations, from its emergence in sources in the mid-15th century up until its extinction in the male line in 1603. All three men – Gaspar (Gašpar) the Elder, Gaspar the Younger, and Matthew (Mate) – had primarily military careers, leading cavalry units and fighting either the Turks or other Christian nobles in civil wars which burdened Croatia, Slavonia, Hungary, and Transylvania from the late 15th to the early 17th century. Gaspar the Elder was the vice-ban of Croatia-Dalmatia and is a relatively well-known figure in Croatian historiography, while the lives of his son and grandson are thoroughly researched for the first time in this article. Gaspar the Younger, initially a supporter of the Habsburgs, was fighting the Ottomans in Croatia until 1532, with significant success, and was later engaged in civil strife in Slavonia, changing the sides he supported several times. He finally opted for King John Zápolya around 1538 and migrated to Zápolya’s realm, settling finally in Transylvania, where he gained many estates and served several de jure and de facto rulers, including another fellow Croat – the bishop of Oradea, George Martinuzzi (Juraj Utišenović Martinušević). His son Matthew, the last male member of this line of the Perušić family, spent his lifetime as a military commander for various Transylvanian rulers, almost always joining the winning side in the conflict and gaining the house in the informal capital – Alba Iulia. He died in a battle in 1603, survived by his sisters’ (Catherine’s and Anna’s) descendants.

Lifespan-Associated Gene Expression Signatures of Recombinant BXD Mice Implicates Coro7 and Set in Longevity.

Although genetic approaches have identified key genes and pathways that promote longevity, systems-level approaches are less utilized. Here, we took advantage of the wealth of omics data characterizing the BXD family of mice. We associated transcript and peptide levels across five tissues from both female and male BXD isogenic lines with their median lifespan. We identified over 5000 genes that showed a longevity correlation in a given tissue. Surprisingly, we found less than 1% overlap among longevity-correlating genes across tissues and sex. These 1% shared genes consist of 51 genes, of which 13 have been shown to alter lifespan. Only two genes -Coro7 and Set- showed a longevity correlation in all tissues and in both sexes. While differential regulation of aging across tissues and sex has been reported, our systems-level analysis reveals two unique genes that may promote healthy aging in unique sex- and tissue-agnostic manner.

Comparative Staging of Blastoderm Development at Oviposition in Eggs from Layer and Broiler Grandparent Breeder Flocks

Recent decades were characterized by genetic selection of broiler and layer chickens for enhanced growth rate and meat yield or intensified egg production, respectively. It is to be expected that genetic selection for various traits would also influence embryo development. The effects of different strains of chickens on egg weight, stage of blastoderm, variation of the blastoderm development and Pearson correlation between egg weight and stage of blastoderm at oviposition were studied in the current study. Hatching eggs were randomly collected from three layer chicken lines (Layer-1, Layer-2, and Layer-3) raised in Poultry Research Institute, and two broiler grandparent flocks (Female and Male) from Aviagen Anadolu. The age of all chicken lines was 36 week. A total of 30 eggs in each line were used, and egg weight and the blastoderm development of the embryos were determined at oviposition time. At the end of the research, significant difference in egg weights (P0.05). The mean stage of blastoderm development in male grandparent line eggs significantly lower than other four genetic lines (P

The New Genealogical Tree of the Da Vinci Family for Leonardo’s DNA. Ancestors and descendants in direct male line down to the present XXI generation

The New Genealogical Tree of the Da Vinci Family for Leonardo’s DNA. Ancestors and descendants in direct male line down to the present XXI generation

Abstract 2256: A population-based mouse model for an experimental basal-like mammary cancer

Abstract The prevalence of obesity, an established risk and prognostic factor for basal-like breast cancer (BLBC), has risen dramatically in the US and many other countries over the past 4 decades. Obesity is also associated with increased resistance to several classes of cancer chemotherapeutic agents used for advanced BLBC. Unfortunately the mechanisms underlying chemotherapy resistance, and the inter-individual differences driving resistance, are poorly understood. Population-based experimental mouse models for determining inter-individual genetic differences in chemotherapy pharmacokinetics and treatment-related efficacy or resistance in mammary gland cancer are not well-defined. Our aim is to develop a population-based experimental model based on outcrosses between 30 different Collaborative Cross line females and hemizygous male B6.FVB-Tg(C3-1-TAg) (greater than N15) congenic mice to produce 30 Collaborative Cross (CC) recombinant inbred line intercross (CC RIX) mouse lines to investigate the interactive effects between diet-induced obesity (DIO) and genetic susceptibility to chemotherapy in a genetically heterogeneous mouse model of BLBC. We report here that thirty genetically-different CCRIL X hemizygous B6.FVB-Tg(C3-1-TAg) congenic female and male CC RIX mice observed longitudinally for occurrence of basal-like mammary tumors showed significant differences in both age at first mammary tumor observation and the fraction of females and males in each RIX bearing tumors. Litter size and the number of females and males of each RIX varied across the 30 CC RIX line. The study was well-powered, with at least X males and Y females for each CC RIX line evaluated longitudinally. We observed that 25 of 30 female CC RIX lines presented with BLMC, with distinct times-to-tumor with each CC RIX. The penetrance of the fraction of each CC RIX line (birth cohort) presenting with BLMC varied greatly in females (25-100%). Surprisingly, 11 of 30 male CC RIX lines presented with BLMC, with penetrance of BLMC varying in males between 10-75%. In conclusion, we present preliminary data to support development of an experimental mouse model for BLBC based on the random mating of 8 genetically-diverse homozygous inbred founder lines followed by in-breeding to greater than 95% homozygosity carrying 45 million single-nucleotide and structural variants. Studies are underway using these CC RIX lines to evaluate the genetic underpinnings of obesity-associated resistance to carboplatin in C3-TAg-driven mammary tumors. Citation Format: John Edgar French, William Pressel, Jody Albright, Melissa VerHague, Stephen D. Hursting. A population-based mouse model for an experimental basal-like mammary cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2256.

Marker-assisted breeding of onion (<i>A. cepa L.</i>) maintainer line resistant to Downy mildew

Relevance. F1 hybrids breeding programs of onion are based on the use of nuclear cytoplasmic male sterility (NCMS). Breeding and seed production based on NCMS suggest a development of a female sterile maternal line, a maintainer line and a paternal component. The development of an isogenic pair of sterile line – maintainer line is one of the most labor-, time-consuming and intellectually-intensive stages of a breeding program, the implementation of which becomes more complicated with an increase in the number of traits / genes for which selection is carried out.Methods and results. This paper presents a genetic scheme for the development of a Downy mildewresistant (pathogen P. destructor) maintainer line of onion using marker-assisted selection (MAS). F2 populations were obtained from the cross of the female maintainer inbred line, Bn1- (13) (genotype – cytN msms pdpd) and male inbred line resistant to Downy mildew, No. 136 (genotype – cytT MsMs PdPd). DNA marker DMR1 were used for Downy mildew resistance gene Pd, jnurf13 – for maintainer gene ms and marker system 5`cob: orfA501 – for N, S and T cytoplasms. It was shown that the MAS application allow to develop a stable Downy mildew resistant maintainer line cytN msms PdPd in two generations.

Phenotyping Anther Extrusion of Wheat Using Image Analysis

Phenotyping wheat (Triticum aestivum L.) is time-consuming and new methods are necessary to decrease labor. To develop a heterotic pool of male wheat lines for hybrid breeding, there must be an efficient way to measure both anther extrusion and the size of anthers. Five hundred and ninety-four soft red winter wheat lines in two replications of randomized complete block design were phenotyped for anther extrusion, a key trait for hybrid wheat production. A device was constructed to capture images using a mobile device. Four heads were sampled per line when anthesis was evident for half the heads in the plot. The extruded anthers were scraped onto a surface, their image was captured, and the area of the anthers was taken via ImageJ. The number of anthers extruded was estimated by counting the number of anthers per image and dividing by the number of heads sampled. The area per anther was taken by dividing the area of anthers per spike by the number of anthers per spike. A significant correlation (R=0.9, p<0.0001) was observed between the area of anthers per spike and the number of anthers per spike. This method is proposed as a substitute for field ratings of anther extrusion and to quantitatively measure the size of anthers.

Transcriptome Profiling Reveals Molecular Changes during Flower Development between Male Sterile and Fertile Chinese Cabbage (Brassicarapa ssp. pekinensis) Lines.

Male sterility exists widely in flowering plants and is used as a fascinating tool by breeders for creating hybrid varieties. Herein, stamen samples from male sterile CCR20000 and male fertile CCR20001 lines during two developmental stages were employed to elucidate the molecular changes during flower development in fertile and sterile Chinese cabbage lines. RNA-seq revealed weak transcriptional activity in the sterile line, which may have led to the abnormal stamen development. The differentially expressed genes were enriched in plant hormone, carbon metabolism, and biosynthesis of amino acid pathways. Important genes with opposite patterns of regulation between the two lines have been associated with the male sterility trait. Members of the transcription factor families such as AP2, MYB, bHLH, and WRKY were highly active in the regulation of structural genes involved in pollen fertility. This study generated important genomic information to support the exploitation of the male sterility trait in Chinese cabbage breeding programs.

Generation of 20 human induced pluripotent stem cell lines from patients with focal segmental glomerulosclerosis (FSGS)

Focal segmental glomerulosclerosis (FSGS) is a major cause of familial nephrotic syndrome. We generated 20 induced pluripotent stem cell lines from patients diagnosed with FSGS. The iPSC lines include 8 female and 12 male lines and cover a donor age range from 31 to 78. The lines were generated from peripheral blood mononuclear cells by integration-free reprogramming using Sendai virus vectors. Cell lines were fully characterized regarding their pluripotency and differentiation potential, and quality controlled for karyotypic integrity, identity and clearance of reprogramming vectors. The generated cell lines represent a valuable tool for disease modelling and drug development for FSGS.

Tamoxifen as a Zika Virus Therapeutic

Zika virus (ZIKV) is an arbovirus belonging to the flaviviridae family with a risk assessment that has increased in recent years and was labeled a global health emergency by the WHO in 2016. There are currently no FDA approved treatment options available for ZIKV, so expeditious development of treatment options is urgent. An on-market drug, tamoxifen (Tam), was selected as a promising candidate for repurposing due to its wide range of biological activities and because it has already been shown to possess activity against hepatitus C (HCV), a related flavivirus. Anti-ZIKV activity of Tam was assessed by compound screens using infectious virus and mechanistic details were gleaned from time of addition and virucidal studies. Tam and the active form, 4-hydroxytamoxifen, showed promising antiviral activity in initial compound screening and up to 8 hours post infection, though the virucidal assay indicated that they do not possess any direct virucidal activity. Additionally, Tam was assessed for its activity against ZIKV in the human male germ cell line, SEM-1, and virus titers show diminished replication of ZIKV over a seven day period compared to controls. These data indicate that Tam has potential to be repurposed as an anti-ZIKV therapeutic and warrants further investigation.

Non-viral derivation of a transgene-free induced pluripotent stem cell line from a male beagle dog

We previously reported the non-viral derivation of transgene-free induced pluripotent stem cells (iPSCs) from somatic fibroblasts of a female beagle dog using an optimized induction medium and integration-free episomal vectors. Here, we report novel derivation of a male canine iPSC line OF35Y-iPS, which showed standard characteristics of pluripotency such as a strong gene expression profile of pluripotency markers, differentiation potential into all three germ layers, and normal karyotype (78XY). Furthermore, we demonstrated targeted integration of 2A-EGFP into the canine NANOS3 locus. The novel iPSC line would be a useful resource for stem cell research and regenerative veterinary medicine.

Successful cryopreservation and regeneration of a partridge colored Hungarian native chicken breed using primordial germ cells

Primordial germ cells (PGCs) are the precursors of germline cells that generate sperm and ova in adults. Thus, they are promising tools for gene editing and genetic preservation, especially in avian species. In this study, we established stable male and female PGC lines from 6Hungarian indigenous chicken breeds with derivation rates ranging from 37.5 to 50 percent. We characterized the PGCs for expression of the germ cell-specific markers during prolonged culture in vitro. An in vivo colonization test was performed on PGCs from four Hungarian chicken breeds and the colonization rates were between 76 and 100%. Cryopreserved PGCs of the donor breed (Partridge color Hungarian) were injected into Black Transylvanian Naked Neck host embryos to form chimeric progeny that, after backcrossing, would permit reconstitution of the donor breed. For 24 presumptive chimeras 13 were male and 11 were female. In the course of backcrossing, 340 chicks were hatched and 17 of them (5%) were pure Partridge colored. Based on the backcrossing 1 hen and 3 roosters of the 24 presumptive chimeras (16.6%) have proven to be germline chimeras. Therefore, it was proven that the original breed can be recovered from primordial germ cells which are stored in the gene bank. To our knowledge, our study is a first that applied feeder free culturing conditions for both male and female cell lines successfully and used multiple indigenous chicken breeds to create a gene bank representing a region (Carpathian Basin).

Brd4-bound enhancers drive cell-intrinsic sex differences in glioblastoma

Sex can be an important determinant of cancer phenotype, and exploring sex-biased tumor biology holds promise for identifying novel therapeutic targets and new approaches to cancer treatment. In an established isogenic murine model of glioblastoma (GBM), we discovered correlated transcriptome-wide sex differences in gene expression, H3K27ac marks, large Brd4-bound enhancer usage, and Brd4 localization to Myc and p53 genomic binding sites. These sex-biased gene expression patterns were also evident in human glioblastoma stem cells (GSCs). These observations led us to hypothesize that Brd4-bound enhancers might underlie sex differences in stem cell function and tumorigenicity in GBM. We found that male and female GBM cells exhibited sex-specific responses to pharmacological or genetic inhibition of Brd4. Brd4 knockdown or pharmacologic inhibition decreased male GBM cell clonogenicity and in vivo tumorigenesis while increasing both in female GBM cells. These results were validated in male and female patient-derived GBM cell lines. Furthermore, analysis of the Cancer Therapeutic Response Portal of human GBM samples segregated by sex revealed that male GBM cells are significantly more sensitive to BET (bromodomain and extraterminal) inhibitors than are female cells. Thus, Brd4 activity is revealed to drive sex differences in stem cell and tumorigenic phenotypes, which can be abrogated by sex-specific responses to BET inhibition. This has important implications for the clinical evaluation and use of BET inhibitors.

Oil content determination on sunflower seeds in drought conditions

Sunflower (Helianthus annuus L.) is one of the most important oil crops in the world and mostly produces oil. Due to the global warming and unexpected climatic changes especially extreme heat and droughts in recent years, as a spring crop sunflower affects severely by these climatic conditions in the vegetation period and loses seed yield hardly. Plant breeding longer process taking at least ten years to develop cultivars. Therefore, sunflower breeders should consider future climatic conditions, hence it seems that drought tolerance should consider primarily in their breeding programs. Oil content is the most important trait in sunflower than the major goal in the breeding studies; likewise, it is the most affected yield trait in sunflower from drought conditions. The study was conducted to determine the oil contents from drought some inbred lines and their hybrids developed lately in National Sunflower Project conducting by Trakya Agriculture Research Institute (TARI). Based on the study results; the significant relationships were observed between oil content and other yield traits in drought conditions. Oil content of male lines exhibited more variability than female lines changed as 13.16%–54.05% male lines and 19.12%–52.46% in female lines, and 30.17%–53.68% in the hybrids. While oil content of inbred lines affected negatively mostly from thousand seed weight and head diameter, leaf number, leaf area, and seed yield followed this trait, respectively. Based on path analysis for the contribution of yield traits on determining of oil content; it determined that seed and oil yield were the most directly affected traits then followed by oil yield in sunflower inbred lines in a negative way, and other yield traits mostly affected oil content determination in drought stress conditions over these two traits. The seed and oil yield, leaf area, and thousand seed weight reduced about over 90% in the drought in inbred lines. Severe droughts resulted heavy decreases in sunflower oil content as well as in the seed and oil yield and other yield traits so no need to focus on increasing oil contents over 40%–43% and it should be sacrified from this trait in that kind of conditions. The sunflower hybrid evaluations in three different locations mentioned that thousand seed weight, seed yield, and leaf area as well as leaf number are the most important yield traits affecting positively of oil content determination in drought stress conditions. Practical applications Sunflower is commonly influenced by the environmental conditions, such as hot temperatures and drought as being a spring growing crop. These environmental stresses mostly appear during mid-summer then sunflower suffers from these stressess resulting of seed and oil yield losses with lower oil content. Especially the stress in the grain filling period results in lower oil content in the sunflower growing season. The study results mentioned that sunflower breeders should consider more male inbred lines to develop better oil content hybrids for drought stress. Furthermore, thousand seed weight is also the most important seed characteristics other than seed and oil yield for consisting of oil content in sunflower. Therefore, we have to focus on developing higher seed yield instead of higher oil content because sunflower sacrifice from seed yield to give higher oil content over 40%–43% during the stress conditions.

An assessment of genetic diversity and combining ability of elite castor genotypes suitable for rainfed conditions

A study was conducted to assess the diversity of 72 castor genotypes used as male lines in the hybrid breeding programme in India using multivariate analysis in addition to morphological characterization using DUS (Distinctiveness, Uniformity and Stability) testing guidelines of castor. Fifteen agronomical and three morphological traits were measured in 72 genotypes in field studies conducted for two years. The multivariate data set was subjected to genetic divergence analysis using Mahalonobis’ D2 static and clustered into nine subgroups following Tocher’s method in addition to Principal Component Analysis (PCA). The genotypes from Clusters 1 and 5 and Clusters 2 and 5 are genetically diverse with 556.9 and 469.7 units while cluster 7 genotypes with higher mean seed yield are potential candidates as varieties after multilocation testing. In PCA, the first two canonical variants were significant and accounted for 68 per cent of the inter accession variability. Grouping of castor genotypes was also done based on four grouping characteristics of DUS guidelines like stem color, bloom, capsule spines and maturity. Combining ability analysis indicated that DCS-96, DCS-94, DCS-104, DCS-97 were good general combiners for seed yield while M-568 x DCS-96 yielded 21 per cent standard heterosis over the best check DCH-177.