Wool Follicles Research Articles

Seasonal variation of fibre follicle activity and wool growth in fat-tailed Sanjabi sheep in west Iran.

This experiment was conducted to investigate the seasonal pattern of hair follicle activity, wool growth and fibre diameter (FD) in Sanjabi sheep in west Iran, Kermanshah (34° 18' N and 47° 3' E, elevation 1420 m). Ten male and 10 female Sanjabi sheep with an initial live weight of 32.1 ± 1.3 and 32.7 ± 1.5 (means ± SD), respectively, were used in a 365-day study. A diet was offered with an estimated concentration of 2.18 Mcal metabolizable energy and 130.0 g/kg DM crude protein. Body weight, average daily gain (ADG) and dry matter intake (DMI) were recorded weekly. The percentages of active primary and secondary wool follicles (PAP and PAS), follicle density and the ratio of secondary to primary follicles (S/P) were determined from skin biopsies, taken from the right mid-side of the sheep at monthly intervals. Raw and clean fibre growth rates and FD were measured from left mid-side patches (10 × 10 cm) harvested at the end of every month. There was a gradual increase in live weight throughout the experiment, while ADG and DMI changed in concert with day length. The greatest values for PAP and PAS were observed in summer, whereas lowest were obtained in winter (p < 0.001). Clean wool growth rate and FD were greatest (p < 0.001) in summer and lowest (p < 0.001) in winter. It is concluded that a seasonal cycle of feed intake, body growth, fibre follicle activity, wool growth and FD occur in fat-tailed Sanjabi sheep.

De novo assembly and characterization of skin transcriptome using RNAseq in sheep (Ovis aries)

Wool is produced via synthetic processes of wool follicles, which are embedded in the skin of sheep. The development of new-generation sequencing and RNA sequencing provides new approaches that may elucidate the molecular regulation mechanism of wool follicle development and facilitate enhanced selection for wool traits through gene-assisted selection or targeted gene manipulation. We performed de novo transcriptome sequencing of skin using the Illumina Hiseq 2000 sequencing system in sheep (Ovis aries). Transcriptome de novo assembly was carried out via short-read assembly programs, including SOAPdenovo and ESTScan. The protein function, clusters of orthologous group function, gene ontology function, metabolic pathway analysis, and protein coding region prediction of unigenes were annotated by BLASTx, BLAST2GO, and ESTScan. More than 26,266,670 clean reads were collected and assembled into 79,741 unigene sequences, with a final assembly length of 35,447,962 nucleotides. A total of 22,164 unigenes were annotated, accounting for 36.27% of the total number of unigenes, which were divided into 25 classes belonging to 218 signaling pathways. Among them, there were 17 signal paths related to hair follicle development. Based on mass sequencing data of sheepskin obtained by RNA-Seq, many unigenes were identified and annotated, which provides an excellent platform for future sheep genetic and functional genomic research. The data could be used for improving wool quality and as a model for human hair follicle development or disease prevention.

Biological defleecing: intravenous infusion of amino acid mixtures lacking lysine and methionine creates a weakened zone in the wool staple, which is amenable to mechanical wool harvesting

Conventional shearing of sheep is labour-intensive, expensive and presents significant occupational health and safety risks. The only alternative at present is based on injection of epidermal growth factor, which severs the fibre at the follicle level. This technology cannot be used in pregnant animals and requires application of a net to retain the severed fleece. An alternative is to create a weakened zone within the wool staple, which would be sufficiently strong to retain the fleece on the sheep while a protective covering regrows, but sufficiently weak as to allow painless and automated removal of the fleece. We demonstrate that this approach is possible using mixtures of amino acids lacking lysine and methionine. Initially we demonstrate the relationships between staple strength, a subjective ‘harvestability’ score and a subjective ‘pain’ score, using fleeces from animals treated with varying levels of cortisol to create a wide range of strengths of wool attachment. We assigned a score to the ease with which we could manually break the staples, and also to the animal’s response to breaking the staples still attached to the skin. The relationships between these variables indicated that a staple was considered harvestable and could be removed with minimal skin flinch response at a staple strength of ~10–13 N/kTex. Staples within this range were then produced by intravenous infusion of mixtures of amino acids lacking in lysine and methionine for a 5-day period. The weak point was uniformly created across the entire fleece and when a prototype roller-pin device was applied to the weakened wool, it uniformly broke the fleece of the three sheep tested. The mode of action of the amino acid treatment on wool growth was studied. There was no effect of unbalanced amino acids on the rate of follicle bulb cell division, the number of active wool follicles, or the length of the keratinisation zone in the wool follicle. Fibre diameter was reduced by ~4 microns by treatment, and intrinsic fibre strength (strength relative to cross-sectional area of the wool fibres), was reduced by ~50%. Results of these trials are encouraging but further work is required to develop a practical, on-farm method of altering systemic amino acid supply and to design an automated, high-throughput system of severing the weakened wool.

Differential expression of genes and proteins associated with wool follicle cycling.

Sheep are valuable resources for the wool industry. Wool growth of Aohan fine wool sheep has cycled during different seasons in 1 year. Therefore, identifying genes that control wool growth cycling might lead to ways for improving the quality and yield of fine wool. In this study, we employed Agilent sheep gene expression microarray and proteomic technology to compare the gene expression patterns of the body side skins at August and December time points in Aohan fine wool sheep (a Chinese indigenous breed). Microarray study revealed that 2,223 transcripts were differentially expressed, including 1,162 up-regulated and 1,061 down-regulated transcripts, comparing body side skin at the August time point to the December one (A/D) in Aohan fine wool sheep. Then seven differentially expressed genes were selected to validated the reliability of the gene chip data. The majority of the genes possibly related to follicle development and wool growth could be assigned into the categories including regulation of receptor binding, extracellular region, protein binding and extracellular space. Proteomic study revealed that 84 protein spots showed significant differences in expression levels. Of the 84, 63 protein spots were upregulated and 21 were downregulated in A/D. Finally, 55 protein points were determined through MALDI-TOF/MS analyses. Furthermore, the regulation mechanism of hair follicle might resemble that of fetation.

Identification of microRNAs in Wool Follicles during Anagen, Catagen, and Telogen Phases in Tibetan Sheep

BackgroundWool quality is one of the most important economic traits in sheep. The wool fiber is derived from specialized skin cells that are referred to as wool follicles. To understand the roles of microRNAs (miRNAs) in wool fiber growth, we detected the expression patterns of miRNAs in wool follicles at the anagen, catagen, and telogen stages from Tibetan sheep through Solexa sequencing.ResultsA total of 244 mature miRNAs were identified. Of these, only five miRNAs are listed in the database of sheep miRNAs (miRBase Database V19), and the other 239 miRNAs have not been previously described in this species. Further analyses indicated that 204 miRNAs are evolutionarily conserved among mammal species, whereas 35 of the identified miRNAs were first found specifically in sheep. The expression pattern analyses showed that the expression levels of 39, 34, and 20 of the miRNAs significantly change between anagen and catagen, between anagen and telogen, and between catagen and telogen, respectively. The results of the bioinformatics analysis show that these differentially expressed miRNAs might regulate wool follicle development by targeting genes in many different pathways, such as the MAPK and Wnt pathways, as well as the pathways that regulate the actin cytoskeleton, focal adhesion, and tight junctions. Furthermore, we identified six differentially expressed miRNAs (oar-miR-103-3P, oar-miR-148b-3P, oar-miR-320-3P, oar-miR-31-5P, oar-novel-1-5P, and oar-novel-2-3P) that might target the key genes of the Wnt pathway. It has been reported that the Wnt pathway is critical for wool follicle development. Therefore, these miRNAs may regulate wool development through the Wnt pathway.ConclusionsOur results provide new information on the identification and expression pattern of miRNAs in wool follicles. Our data might therefore aid in the understanding of the mechanisms of wool follicle development in sheep.

Evidence that Notch and Delta expressions have a role in dermal condensate aggregation during wool follicle initiation

Notch pathway genes have been implicated in the commitment of mesenchymal cells to a wool follicle cell fate. Notch1 and Delta1 transcripts were quantified in fetal skin of fine-woolled (Merino) and strong-woolled (Tukidale) sheep at two time points: either preceding (d56) or during (d70) the first wave of follicle initiation. DIG-labelled probes for both transcripts were localised in the epithelium, some mesenchymal cells, and in the dermal condensates of primordia. The possibility that condensates selectively incorporated Delta1-labelled mesenchymal cells is considered. The involvement of Notch1 in condensate formation was also explored in cultured fetal skin explants and whisker papilla cells using DAPT to block Notch signalling. In its presence, follicle initiation in skin explants was reduced, and the propensity for cultured papilla cells to aggregate was abolished. Results suggest that Notch1 activation is a prerequisite for mesenchymal aggregation. It is speculated that Delta interactions contribute to condensate formation, in vivo.

Transient treatment of pregnant Merino ewes with modulators of cortisol biosynthesis coinciding with primary wool follicle initiation alters lifetime wool growth

The economically important characteristics of the adult fleece of Merino sheep, such as increases in clean fleece weight, fibre length, fibre diameter and crimp characteristics are determined during critical phases of fetal development of the skin and its appendages. Genetics plays a major role in the development of traits, but the maternal uterine environment could also influence development. Treatment of pregnant ewes with cortisol and its analogues has previously been shown to produce changes in wool follicle morphology. The aim of this study was to determine the effect of transient manipulation of maternal cortisol status during critical phases of wool follicle initiation and development in utero. From Days 55–65 post-conception, singleton-bearing Merino ewes were treated with metyrapone (cortisol inhibitor) or betamethasone (cortisol analogue). Lambs exposed to metyrapone in utero were born with hairier birthcoats than the control or betamethasone treatment groups (P &lt; 0.05), displayed a 10% increase in staple length and a reduction in crimp frequency for the first three shearings (P &lt; 0.05). Co-expression network analysis of microarray data revealed up-regulation of members of the transforming growth factor-β and chemokine receptor superfamilies, gene families known to influence hair and skin development. These experiments demonstrate that presumptive transient manipulation of maternal cortisol status coinciding with the initiation of fetal wool follicle development results in long-term alteration in fleece characteristics, namely fibre length and fibre crimp frequency. These results indicate it is possible to alter the lifetime wool production of Merino sheep with therapeutics targeted to gene expression during key windows of development in utero.

Cloning and expression in follicle anagen of &lt;I&gt;ILK&lt;/I&gt; gene in sheep

Integrin linked kinase (ILK) is a scaffold protein, which plays important roles in hair follicle development. The cDNA sequence of novel ILK gene in sheep was cloned by PCR method and analyzed by bioinformatics. Tissue expression profiling in eight tissues and temporal profiling at different wool follicle anagen stages in skin was analyzed. The results showed that the whole open reading frame (ORF) of ILK gene was 1 359 bp in length, which encoded 452 amino acids. Bioinformatic analysis indicated that the secondary structure of ILK gene was mainly made up of three ankyrin repeats and a kinase domain, and there were multiple phosphorylation and Protein Kinase C sites in this gene. The RT-PCR result confirmed that ILK mRNA was expressed in heart, liver, spleen, lung, skeletal muscle, skin, and small intestine, and the expression level was much higher in skin, spleen, and liver than others. The q-PCR analysis demonstrated that the ex-pression level of ILK was significantly increased from March to May (early follicle anagen initiation) in both sheep breeds, Chinese Merino and Kazakh sheep, and there were certain differences from June to October between the two breeds. The above results indicated that ILK gene may play key roles in regulating secondary follicle growth.

The germinative epithelium of sheep vibrissae and wool follicles has extensive proliferative potential but is dependent on the dermal papilla

Aim:To investigate the growth potential of keratinocytes derived from the germinative epithelium (GE) of ovine hair follicles. Stem cells from the outer root sheath (ORS) of hair follicles migrate to the GE in the lower follicle where they proliferate and differentiate to form the hair fiber. It has been suggested that the GE comprises transit-amplifying cells and that the duration of anagen is determined by their limited proliferative potential. However, we show here that keratinocytes derived from the GE of ovine follicles grow extensively in vitro, arguing against this hypothesis.Materials and Methods:Primary cultures of keratinocytes were initiated from microdissected GE tissue from ovine vibrissae and wool follicles. Clonal lines of keratinocytes were derived by limiting dilution. Their growth potential was determined by exhaustive serial passaging. Expression of differentiation markers was evaluated by real-time polymerase chain reaction.Results:Initiation of these cultures required that interaction between the GE and dermal papilla was maintained. However, the keratinocytes could subsequently be cloned and were grown as pure cell populations for 26-52 cell doublings. This proliferative potential is several orders of magnitude greater than required to maintain a single anagen phase. The keratinocytes were indistinguishable from ORS keratinocytes from the same follicles, expressing K14 while undifferentiated, and upregulating the epidermal and inner root sheath markers, loricrin and KRT27 on differentiation. Thus, these cells initially depend on papilla-derived signals to grow, but can revert to an ORS-like phenotype in vitro. Their extensive proliferative capacity shows that the GE is not an exclusively transit-amplifying cell population.

Search for Variation in the Ovine KAP7-1 and KAP8-1 Genes Using Polymerase Chain Reaction–Single-Stranded Conformational Polymorphism Screening

Keratins and keratin-associated proteins (KAPs) are large heterogeneous groups of proteins that constitute about 90% of the wool fiber. The genes encoding the high glycine-tyrosine (HGT) KAPs are the first sub-group of KAP genes expressed in the wool follicle and just after expression of the keratin genes. Little is known about variation in these genes, which led us to investigate two HGT-KAP genes, KRTAP7-1 and KRTAP8-1. Polymerase chain reaction-single-stranded conformational polymorphism analysis was used to investigate these genes in 250 Romney-cross sheep. For KRTAP7-1, two unique banding patterns were detected for amplicons that spanned the entire coding region. Sequencing confirmed the presence of two sequences with only one nucleotide difference (c.173G/A) putatively resulting in p.Ser58Asn. One was identical to the published ovine KRTAP7-1 sequence. For KRTAP8-1, five unique banding patterns were detected in an amplicon that spanned the entire coding region. Sequencing revealed five different DNA sequences, all of which were highly homologous to the previously reported ovine KRTAP8-1 sequence. Among these five sequences, four single-nucleotide substitutions were identified and three of them were located in the coding region. One of these was nonsynonymous and would putatively result in p.Tyr34Asn. The variation detected in KRTAP7-1 and KRTAP8-1 may influence their expression or protein structure.

Determination of Wool Follicle Characteristics of Iranian Sheep Breeds

In this study wool follicle characteristics of Iranian sheep breeds were investigated. A total of 242 male and female sheep of different age from 5 breeds were studied. Samples of skin were taken from the right midside to determine follicle characteristics. Overall averages for males and females were 3.7±0.1 and 3.5±0.1 for S/P ratio; 3.2±0.1 and 3.6±0.1 for primary follicle density; 12.1±0.7 and 13.8±0.4 for secondary follicle density; 15.3±0.7 and 17.4±0.4 for total primary plus secondary follicle density; 2.3±0.2 and 3.5±0.3 for percentage of inactive secondary follicles. Significant differences were found in some follicle characteristics between sheep breeds. Results of this study showed that sheep breeds raised in different parts of Iran characterized by a low S/P ratio

Maternal nutrition during pregnancy influences offspring wool production and wool follicle development1

The effects of maternal nutrition on offspring wool production (quality and quantity) were evaluated. Primiparous Rambouillet ewes (n = 84) were randomly allocated to 1 of 6 treatments in a 2 × 3 factorial design. Selenium treatment [adequate Se (ASe, 9.5 μg/kg of BW) vs. high Se (HSe, 81.8 μg/kg of BW)] was initiated at breeding, and maternal nutritional intake [control (CON, 100% of requirements) vs. restricted (60% of CON) vs. overfed (140% of CON)] was initiated at d 50 of gestation. Lamb birth weight was recorded at delivery, and all lambs were placed on the same diet immediately after birth to determine the effects of prenatal nutrition on postnatal wool production and follicle development. At 180 ± 2.2 d of age, lambs were necropsied and pelt weights were recorded. Wool samples were collected from the side and britch areas, whereas skin samples were collected from the side of each lamb only. Although Se status did not influence side staple length in males, female lambs born from ewes on the ASe treatment had a shorter staple length (P < 0.05) when compared with females from ewes on the HSe treatment. Maternal nutritional intake and Se status did not influence (P ≥ 0.23) wool characteristics on the britch. However, at the britch, wool from female lambs had a reduced comfort factor (P = 0.01) and a greater (P = 0.02) fiber diameter compared with wool from male lambs. Maternal Se supplementation, maternal nutritional plane, sex of the offspring, or their interactions had no effect (P > 0.13) on primary (29.10 ± 1.40/100 µm(2)) and secondary (529.84 ± 21.57/100 µm(2)) wool follicle numbers. Lambs from ASe ewes had a greater (P = 0.03) secondary:primary wool follicle ratio compared with lambs from HSe ewes (20.93 vs. 18.01 ± 1.00). Despite similar postnatal diets, wool quality was affected by maternal Se status and the maternal nutritional plane.

The effects of short-term manipulation of thyroid hormone status coinciding with primary wool follicle development on fleece characteristics in Merino sheep

Thyroidectomy surgery performed late in gestation results in perturbations in wool follicle development in foetal sheep, showing the importance of thyroid hormones for wool follicle development. The aim of this study was to determine the influence of transient manipulation of thyroid hormone status at a time corresponding with foetal primary wool follicle initiation. Pregnant Merino ewes (n = 12 per treatment) were treated daily between gestational days 55 and 64 with control (vehicle), exogenous thyroxine (T4) or propylthiouracil (PTU), an inhibitor of T4 synthesis, and conversion to the active form of the thyroid hormone (triiodothyronine). There were no significant differences in birth weight, gestational lengths and birth coat scores of the resultant lambs. The total primary and secondary follicle densities were significantly lower in lambs exposed to exogenous T4 compared with other treatments (P < 0.05). However, the T4 group displayed a higher proportion of mature secondary follicles (reflected by increased mature secondary follicle densities and mature secondary/primary follicle ratios) than the other treatment groups (P < 0.05). The skin morphology of the lambs differed 12 months later, with the T4 group having significantly higher total follicle densities compared with the PTU group, largely attributed to increased mature and total secondary follicle densities. However, this increase in wool follicle densities did not translate to differences in the fleece yields and weight, fibre diameter, staple lengths or any other fibre parameters. This study showed that transient manipulation of thyroid hormone status during foetal primary follicle initiation does have long-term consequences on the morphology of wool follicles, in particular the maturity of secondary wool follicles.

Pathophysiology in Teladorsagia (Ostertagia) circumcincta-infected sheep selected for high fleece weight

Resilience to parasitism is considered to be the maintenance of growth and production during infection, probably associated with an immune response with lesser detrimental effects on the host relative to adverse effects on the parasite. Resilience to infection with Teladorsagia circumcincta was investigated in lambs from a flock selected for forty generations for high fleece weight (HFW), but with higher FEC and worm burdens than their unselected control (C) flock run in parallel. After recovery from surgery to implant abomasal cannulae, four parasite-naïve lambs from each flock were infected intraruminally at 6.5 months-of-age with 50,000 T. circumcincta L3, then from Day 35 to 70 post infection with 10,000 larvae at weekly intervals. Blood, abomasal fluid and faecal samples were collected daily to Day 35 and thence twice weekly for measurement of serum gastrin and pepsinogen concentrations, blood eosinophils, abomasal pH and FEC. Abomasal worm counts were made after necropsy on Day 94. Skin biopsies were collected weekly for estimation of the percentage of wool follicles containing paracortical cells. Total serum immunoglobulin and IgG1, IgG2, IgA and IgM titres specific for T. circumcincta antigens were estimated twice weekly to Day 42 p.i., then weekly. After the primary challenge, FEC were higher in the HFW lambs, whereas neither group shed many eggs during the 5-week trickle infection; worm burdens were small at post mortem. Resilient HFW lambs showed a lesser inflammatory response, but relatively small differences in abomasal secretion. Circulating eosinophil counts increased moderately in both groups, less in the HFW lambs, during the primary infection and more markedly during the subsequent trickle infection, when the increase in the C lambs became significantly greater. All measured serum antibody titres were low in both groups throughout. Selection for HFW altered the wool characteristics of parasite-naïve lambs (fewer follicles containing paracortical cells). There was a slower increase in the percentage of follicles containing these cells after primary infection. Abomasal function was similar in the two groups, both exhibiting typical increases in abomasal pH and serum gastrin and pepsinogen concentrations. The most marked differences in the HFW lambs were a greater rise in serum pepsinogen during the primary infection and the 2-day delay in onset of hypoacidity. Resilience to parasitism in this flock is consistent with maintenance of wool quality and small differences in abomasal secretion resulting from an attenuated immune response causing fewer detrimental effects on host tissues.

Programming sheep production on saltbush: adaptations of offspring from ewes that consumed high amounts of salt during pregnancy and early lactation

We investigated if feeding a high salt diet (pellet containing 14% NaCl) or saltbush (Atriplex nummularia) to ewes between day 60 of gestation and day 21 of lactation would allow their offspring to gain more weight, and produce more wool, when grazing saltbush as adults compared to offspring from ewes that were fed a control diet (2% NaCl) or grazed pasture. At 10 months of age, offspring grazed saltbush for 8 weeks then dry pasture for 2 weeks. Throughout this time, liveweights, plasma renin activity and wool growth (g/day) were measured. Greasy and clean fleece weights, and fleece characteristics were measured at 14 months of age, and greasy fleece weight was measured again at 22 months after grazing pasture. Offspring from ewes that consumed the high salt pellet had an 8 and 10% increased fleece weight at 14 and 22 months of age, respectively (P ≤ 0.01). Offspring of ewes that consumed saltbush also showed an 8% increase in greasy fleece weight at 22 months of age (P ≤ 0.05). Offspring from ewes that consumed saltbush had lower plasma renin activity and gained tissue weight when grazing saltbush (P ≤ 0.05), whereas the other three treatment groups all lost weight (P &gt; 0.05). Grazing pregnant ewes on saltbush induces important adaptations in plasma renin activity of their offspring, which allows them to gain weight when grazing saltbush as adults and may also increase the density of their wool follicles. Grazing pregnant ewes on saltbush can profit farmers in three main ways: (i) ability to utilise salt-affected land; (ii) increase weight gain of sheep when grazing saltbush; and (iii) increase fleece weight.

Expression patterns of keratin intermediate filament and keratin associated protein genes in wool follicles

The catalogue of hair keratin intermediate filaments (KIFs) and keratin-associated proteins (KAPs) present in wool follicles is incomplete. The full coding sequences for three novel sheep KIFs ( KRT27, KRT35 and KRT38) and one KAP ( KRTAP4-3) were established in this study. Spatial expression patterns of these and other genes ( KRT31, KRT85, KRTAP6-1 and trichohyalin) were determined by in situ hybridisation in wool follicles at synchronised stages of growth. Transcription proceeded in the order: trichohyalin, KRT27, KRT85, KRT35, KRT31, KRT38, KRTAP6-1 and KRTAP4-3, as determined by increasing distance of their expression zones from the germinal matrix in anagen follicles. Expression became gradually more restricted to the lower follicle during follicle regression (catagen), and ceased during dormancy (telogen). Some genes ( KRT27, KRT31, KRT85 and KRTAP6-1), but not others, were expressed in cortical cells forming the brush-end, indicating specific requirements for the formation of this anchoring structure. The resumption of keratin expression was observed only in later stages of follicle reactivation (proanagen). KIF expression patterns in primary wool follicles showed general resemblance to their human homologues but with some unique features. Consistent differences in localisation between primary and secondary wool follicles were observed. Asymmetrical expression of KRT27, KRT31, KRT35, KRT85 and trichohyalin genes in secondary follicles were associated with bulb deflection and follicle curvature, suggesting a role in the determination of follicle and fibre morphology.

Notch pathway gene expression and wool follicle cell fates

The Notch family of genes has been implicated in specifying cell fates during hair follicle morphogenesis. We examined Notch gene expression during wool follicle formation, as an understanding of genes that influence cell distributions in the developing follicle is a prerequisite for devising molecular strategies to manipulate fibre characters and follicle density. We identified transcripts for the Notch1 receptor and one of its ligands, Jagged1, in fetal sheep skin by reverse transcriptase polymerase chain reaction. The sheep-specific cDNA sequences were used as templates to produce probes to investigate the expression patterns of Notch1 and Jagged1 in developing ovine fetal skin by in situ hybridisation. Notch1 and Jagged1 were detected in the epidermis and in a subpopulation of mesenchymal cells before follicle initiation. At day 70 during follicle initiation, transcripts were also detected in cells at the tip of the epidermal plug and in dermal condensates. By day 86, Notch1 and Jagged1 were detected in the distal cells of the epidermal downgrowths and epidermis and Notch1 was no longer detected in the mesenchyme and dermal condensates. After day 96, transcripts were absent from the epidermis, but localised to differentiating outer root sheath (ORS) cells. The distributions of transcripts implicate a Notch1–Jagged1 signal pathway in the fates of prospective ORS cells. The transient appearance of Notch1 in cells at the epidermal–mesenchymal junction during early follicle morphogenesis suggests that the receptor may be responsible for the specification of a cell subpopulation committed to a prepapilla fate at initiation.

Characterization of transport systems for cysteine, lysine, alanine, and leucine in wool follicles of sheep

Aspects of the uptake of the AA Cys, Leu, Ala, and Lys into wool follicles were investigated using short-term culture of thin strips of sheep skin. Following verification of the reliability of the model system, the sites of uptake of the radiolabeled AA were shown to differ and to be consistent with their different roles in fiber production. Cysteine appeared in the zone of keratinization immediately distal to the follicle bulb. Lysine was incorporated into the germinative cells of the follicle bulb and the cells of the inner root sheath. Leucine and Ala were incorporated into the follicle bulb, inner root sheath, and keratinizing fiber. The incorporation of all AA into the dermal papilla was low. The relative rates of uptake of the AA into the wool follicle were as follows: L-Cys (100), L-Leu (5.5), L-Ala (2.5), and L-Lys (0.8). Uptake of Cys was saturable and followed Michaelis-Menten kinetics, suggesting a carrier-mediated system, with little or no diffusion. The majority (70%) of Cys uptake into follicles was via a Na-independent system that was not inhibited by alpha-(methyl-amino)isobutyric acid or 2-amino-2-norbonanecarboxylic acid and therefore is not via the normal Cys transport systems A, ASC, or L. Uptake of Cys appeared to be via a low-affinity, high-capacity transport system, which may be unique to the fiber-producing follicle. The majority of Ala transport had characteristics consistent with the functioning of system A (Na-dependent, inhibited by alpha-(methylamino)isobutyric acid, and low substrate affinity). Leucine uptake was inhibited by 2-amino-2-norbonanecarboxylic acid but was Na-dependent, suggesting that a variant of system L operates in the follicle to transport Leu. Lysine uptake was consistent with the operation of the usual Lys transporter system y+. Diets designed to maximize wool growth should provide AA profiles reflecting the relative rates of uptake demonstrated in this study. Investigations of possible polymorphisms in genes encoding AA transport proteins in follicles may reveal a source of genetic differences in wool growth potential among genotypes.

Protein synthesis rates in skin components and skeletal muscle of sheep selected for divergent clean fleece weight in response to below- and above-maintenance nutrition

Protein metabolism in skin and muscle was studied in Merino wethers selected for high (F+, n = 10) or low (F–, n = 10) estimated breeding values for clean fleece weight, but with similar estimated breeding values for liveweight and fibre diameter, raised to 20 months of age under the same conditions, and then offered two levels of nutrition (0.8 or 1.8 × maintenance) for 37 days. Over 37 days, F+ sheep had greater rate of wool production, liveweight gain, and had greater eye-muscle and fat depth than F– sheep (P &lt; 0.05). Fractional synthesis rates of protein (%/day) in the epidermis, dermis, whole skin and muscle were affected by both feeding level (P &lt; 0.05) and genotype (P &lt; 0.05). The fractional synthesis rates of protein were greater (P &lt; 0.05) in F+ sheep at both levels of intake. There was an interaction (P &lt; 0.01) between genotype and feeding level for the protein fractional synthesis rate in muscle, where F+ sheep were more responsive to higher feed intake. Muscle of F– sheep responded to increased amino acid supply by reducing the rate of protein degradation without altering synthesis rate; whereas muscle of F+ sheep responded by increasing the rates of both protein synthesis and degradation. The overall muscle fractional synthesis rate (1.6%/day) was ~7-times lower than the skin fractional synthesis rate (10.8%/day) in these animals (P &lt; 0.01). F+ sheep had a higher rate of protein synthesis in dermis and whole skin to support their higher wool protein accretion at both levels of feed intake. Muscle protein synthesis rate was greater in F+ sheep offered above-maintenance metabolisable energy (ME) intake than those given below-maintenance ME intake but was unaffected by ME intake in F– sheep. The results indicate that selection for wool growth not only affects production of wool and the wool follicle, but also affects the rate of protein turnover in components of the skin and skeletal muscle.

Perinatal wool follicle attrition coincides with elevated perinatal circulating cortisol concentration in Merino sheep

The development of the follicle population in Merino sheep skin was studied from 30 days pre-partum to 112 days post-partum in single- and twin-born Merino lambs. The total number of primary follicles estimated from primary follicle density and skin surface area did not change over this period, while secondary follicle number increased to Day 28 post-partum in singles and Day 56 post-partum in twins. Twins had 14% fewer total follicles than singletons (P &lt; 0.001), largely reflecting a bodyweight (hence estimated skin surface area) difference of 10%, with little difference in total follicle density (P &gt; 0.05). Immediately post-partum there was a 36% decrease (P &lt; 0.0001) in the secondary to primary follicle ratio (S/P) of the twin lambs, while a small non-significant decrease (7%; P &gt; 0.05) occurred in the single lambs. This attrition coincided with a surge in plasma cortisol concentration that commenced ~12 days before birth and persisted for 6–12 days post-partum. The surge in plasma cortisol concentration occurred in both single and twin lambs, commencing on Day 138 of gestation; however, the peak cortisol concentration and the period over which cortisol remained elevated was greater in twin lambs than in singletons (P &lt; 0.001). This study confirms a previous finding of a perinatal reduction in S/P ratio in Merino sheep. The reduction was greater in twin lambs than in singletons but the follicles of twins recovered rapidly so that there was little difference in final S/P ratio between birth types after Day 30 post-partum. The postnatal follicle reduction followed the perinatal surge in plasma cortisol concentration and appeared to reflect the magnitude of the cortisol surge in that twins, which displayed a higher peak cortisol concentration, had a greater reduction in active follicles than singletons.