Ciliary Morphology Research Articles

Evidence for the involvement of caspases in establishing proper cerebrospinal fluid hydrodynamics

A large number of cells undergo apoptosis via caspase activation during and after neural tube closure (NTC) in mammals. Apoptosis is executed by either intrinsic or extrinsic apoptotic pathways, and inhibition of each pathway causes developmental defects around NTC stages, which hampers the physiological roles of apoptosis and caspases after NTC. We generated transgenic mice in which a broad spectrum of caspases could be suppressed in a spatiotemporal manner by pan-caspase inhibitor protein p35 originating from baculovirus. Mice with nervous system-specific expression of p35 (Nestin-Cre (NCre);p35V mice) exhibited postnatal lethality within 1 month after birth. They were born at the expected Mendelian ratio, but demonstrated severe postnatal growth retardation and hydrocephalus. The flow of cerebrospinal fluid (CSF) between the third and fourth ventricles was disturbed, whereas neither stenosis nor abnormality in ciliary morphology was observed in the pathway of CSF flow. Hydrocephalus and growth retardation of NCre;p35V mice were not rescued by the deletion of RIPK3, an essential factor for necroptosis which occurs in the absence of caspase-8 activation during development. The CSF of NCre;p35V mice contained a larger amount of secreted proteins than that of the controls. These findings suggest that the establishment of proper CSF dynamics requires caspase activity during brain development after NTC.

Sulforaphane attenuates cisplatin-induced hearing loss by inhibiting histone deacetylase expression

IntroductionCruciferous vegetables are a rich source of sulforaphane (SFN), which acts as a natural HDAC inhibitor (HDACi). Our previous study found that HDACi could restore histone acetyltransferase/histone deacetylase (HAT/HDAC) balance in the cochlea and attenuate gentamicin-induced hearing loss in guinea pigs. Here, we investigated the protective effect of SFN on cisplatin-induced hearing loss (CIHL).MethodsThirty rats were randomly divided into 3 equal groups: the control group, cisplatin group, and SFN+cisplatin group. Rats were injected with SFN (30 mg/kg once a day) and cisplatin (7 mg/kg twice a day) for 7 days to investigate the protective role of SFN on CIHL. We observed auditory brainstem response (ABR) threshold shifts and immunostained cochlear basilar membranes of rats. For in vitro experiments, we treated HEI-OC1 cells and rat cochlear organotypic cultures with SFN (5, 10, and 15 μM) and cisplatin (10 μM). Immunofluorescence, cell viability, and protein analysis were performed to further analyze the protective mechanism of SFN on CIHL.ResultsSFN (30 mg/kg once a day) decreased cisplatin (7 mg/kg twice a day)-induced ABR threshold shifts and outer hair cell loss. CCK-8 assay showed that cisplatin (10 μM) reduced the viability of HEI-OC1 cells to 42%, and SFN had a dose-dependent protective effect. In cochlear organotypic cultures, we found that SFN (10 and 15 μM) increased cisplatin (10 μM)-induced myosin 7a+ cell count and restored ciliary morphology. SFN (5, 10, and 15 μM) reversed the cisplatin (10 μM)-induced increase in HDAC2, -4, and -5 and SFN (15 μM) reversed the cisplatin (10 μM)-induced decrease in H3-Ack9 [acetyl-histone H3 (Lys9)] protein expression in HEI-OC1 cells. Neither cisplatin nor cisplatin combined with SFN affected the expression of HDAC7, or HDAC9.ConclusionSFN prevented disruption of the HAT/HDAC balance, protecting against CIHL in rats.

Spontaneous rotation of a toric implantable collamer lens related to abnormal ciliary body morphology: a case report

BackgroundThis is a case of spontaneous toric implantable collamer lens (TICL) rotation that occurred twice in the left eye of a patient.Case presentationA 24-year-old gentleman received TICL implantation for treatment of myopic astigmatism encountered with spontaneous rotation of approximately 90° from its original position. TICL reposition procedure was performed with visual outcome of 20/16. Surprisingly, a re-rotation of TICL occurred. The patient underwent a replacement of non-toric ICL with a larger size after careful re-evaluation with final visual outcome of 20/20. A short and small ciliary process with shallow ciliary sulcus and posteriorly positioned ciliary body was found by Ultrasound Biomicroscopy (UBM).ConclusionsThe unique morphology of the ciliary body may have a potential association with the vault and instability of implanted ICL. Careful examination of the ciliary body morphology is essential in preoperative evaluation.

The novel ciliogenesis regulator DYRK2 governs Hedgehog signaling during mouse embryogenesis.

Mammalian Hedgehog (Hh) signaling plays key roles in embryogenesis and uniquely requires primary cilia. Functional analyses of several ciliogenesis-related genes led to the discovery of the developmental diseases known as ciliopathies. Hence, identification of mammalian factors that regulate ciliogenesis can provide insight into the molecular mechanisms of embryogenesis and ciliopathy. Here, we demonstrate that DYRK2 acts as a novel mammalian ciliogenesis-related protein kinase. Loss of Dyrk2 in mice causes suppression of Hh signaling and results in skeletal abnormalities during in vivo embryogenesis. Deletion of Dyrk2 induces abnormal ciliary morphology and trafficking of Hh pathway components. Mechanistically, transcriptome analyses demonstrate down-regulation of Aurka and other disassembly genes following Dyrk2 deletion. Taken together, the present study demonstrates for the first time that DYRK2 controls ciliogenesis and is necessary for Hh signaling during mammalian development.

The Atoh1 expression levels are correlated with the arrangement, ciliary morphology, and electrophysiological characteristics of ectopic hair cell-like cells

Previous reports have suggested that the level and duration of Atoh1 expression are correlated with the survival, arrangement and stereociliary bundle-related morphology of hair cells during development, but whether Atoh1 expression levels are correlated with the arrangement, bundle formation and electrophysiological characteristics of newly formed hair cells is unknown. To address this question, cultured cochlear explants obtained from neonatal rats were treated with different titers of a human adenovirus serotype 5 (Ad5) vector encoding Atoh1 and/or EGFP (EGFP-Atoh1+/-). The results showed that higher EGFP-Atoh1 concentrations led to higher initial Atoh1 mRNA expression levels and induced greater numbers of ectopic hair cell-like cells (EHCLCs) in the lesser epithelial ridge (LER). Furthermore, gradual increases in the number of EHCLCs were associated with the progressive conversion of the LER region similarly to that of hair cells during development. Some of the cilia on EHCLCs with higher Atoh1 expression were regularly arranged in a manner similar to that of normal hair bundles. As demonstrated through patch clamp recordings, high Atoh1 expression was associated with significantly decreased proportions of cells with Ih currents, significantly reduced proportions of transient potassium channel currents, and potassium channel currents with a greatly increased mean amplitude, which indicated that EHCLCs with high Atoh1 expression were more mature than those with low Atoh1 expression. Overall, the evidence suggests that the Atoh1 expression levels affect not only the arrangement and ciliary morphology of hair cells but also the electrophysiological characteristics of Atoh1-induced EHCLCs, and these findings provide important guidance for future therapies aimed at treating deafness.

Ciliary muscle morphology and accommodative lag in hyperopic anisometropic children.

To investigate the cross-sectional area (CSA) and thickness of the ciliary muscle and their correlation with accommodative lag in hyperopic anisometropic children. Forty children aged between 6 and 10years with hyperopic anisometropia were recruited. The more hyperopic eye (mean refractive power of 3.51 ± 1.70 D) was compared with the less hyperopic eye (mean refractive power of 0.78 ± 1.41 D). The thickness and CSA of the ciliary muscle were measured with anterior segment optical coherence tomography (OCT) images at four meridians. The differences between the eyes and the correlations between CSA, thickness, axial length and accommodative lag were accessed. There was no statistically significant difference in CSA between the two groups at any meridian, except at the inferior meridian (P < 0.05). There was no statistically significant difference in ciliary muscle thickness between eyes at any meridian, except on the temporal and the nasal meridians (P < 0.05). There was a significant difference in the ratio of CSA to axial length at all meridians between the two groups (all P < 0.05). Accommodative lag was 1.65 ± 0.55 D and 0.93 ± 0.45 D in the more and less hyperopic eyes, respectively, which was a statistically significant difference (P < 0.05). There was no significant correlation between the CSA with the axial length and the accommodative lag. This study demonstrated a greater degree of accommodative lag in the more hyperopic eye of anisometropic children. There was no correlation among accommodative lag, axial length and CSA of the ciliary muscle with the degree of hyperopia.

Emmetropes and myopes differ little in their accommodation dynamics but strongly in their ciliary muscle morphology

Previous work suggested an association between near vision and myopia. We therefore investigated the accommodation process in emmetropes and myopes regarding morphologic changes of the ciliary muscle (CM) and power changes of the lens for different accommodation demands. The temporal CM of 18 emmetropic and 20 myopic students was imaged via anterior segment optical coherence tomography during far and near accommodation (2.5D, 3D, 4D). Additionally, accommodation dynamics to the stimuli pattern far-near-far (15 s each; 2.5D, 3D, 4D) were recorded with eccentric infrared photorefraction. OCT images were processed using custom-developed software facilitating the analysis of selective CM thickness (CMT) readings and CMT profiles. Anterior CMT readings were significantly smaller in myopes. Starting at 1.4 mm posterior to the scleral spur (SP), myopic CM became thicker than emmetropic. Anterior CMT changes (ΔCMT) continuously increased with accommodation demand in myopes while emmetropic ΔCMT only increased from 2.5D to 3D. Compared to emmetropes, myopes showed smaller ΔCMT but increased CM movement relative to SP. There were no significant differences between the groups for accommodation changes from far to near vision and vice versa, velocity, microfluctuations, power spectra or lag of accommodation. At 4 D, larger ΔCMT were associated with lower lens changes for disaccommodation. While CM shape, movement, and thickness showed distinct differences depending on refractive error, emmetropes and myopes did not differ in their dynamic accommodation. Further analysis is necessary to evaluate whether the CM’s anatomical shape or predispositions in its intramuscular constituents are causative factors in myopigenesis.

Prolonged nearwork affects the ciliary muscle morphology

There is evidence for a possible link between myopia development and near vision. We investigated the effect of prolonged nearwork on ciliary muscle (CM) morphology and accommodation in 18 myopic and 17 emmetropic subjects (age 19 to 25). The CM was imaged during far (0.25 D) and near vision (4 D) using optical coherence tomography (OCT), and accommodation to a step pulse (0.25 D - 4 D - 0.25 D, 15 s each) was assessed by eccentric infrared photorefraction before and after a 30-min reading task at 25 cm. OCT images were analyzed using a custom-developed semi-automatic segmentation algorithm to determine CM thickness (CMT) profiles and selective CMT readings. Accommodation was assessed using a non-linear model. On average, the CM got thinner after nearwork, predominantly at 0.0–1.4 mm posterior to the scleral spur in emmetropes, and at 1.0–1.9 mm in myopes. Selective CMT readings confirmed a significant thinning after nearwork (univariate ANOVA F1,66 = 26.313, p < 0.001), without any influence of the subjects’ refractive state (F1,66 = 1.887, p = 0.174) or the target distance (F1,66 = 0.014, p = 0.907). The mean accommodation response for targets at infinity was significantly increased after nearwork (F1,32 = 7.775, p = 0.009), with a larger myopic shift in myopes (F1,32 = 11.310, p = 0.002). No change in velocity of accommodation was found. Sharing properties of striated muscles, the CM was expected to increase its thickness, but the opposite was found. Previous studies suggesting sustained nearwork to result in a CM spasm cannot be confirmed by the data presented here. Further research exploring the possible impact of sympathetic innervation is necessary as it is activated during intense nearwork.

Fibroblast growth factor receptor influences primary cilium length through an interaction with intestinal cell kinase

Vertebrate primary cilium is a Hedgehog signaling center but the extent of its involvement in other signaling systems is less well understood. This report delineates a mechanism by which fibroblast growth factor (FGF) controls primary cilia. Employing proteomic approaches to characterize proteins associated with the FGF-receptor, FGFR3, we identified the serine/threonine kinase intestinal cell kinase (ICK) as an FGFR interactor. ICK is involved in ciliogenesis and participates in control of ciliary length. FGF signaling partially abolished ICK's kinase activity, through FGFR-mediated ICK phosphorylation at conserved residue Tyr15, which interfered with optimal ATP binding. Activation of the FGF signaling pathway affected both primary cilia length and function in a manner consistent with cilia effects caused by inhibition of ICK activity. Moreover, knockdown and knockout of ICK rescued the FGF-mediated effect on cilia. We provide conclusive evidence that FGF signaling controls cilia via interaction with ICK.

Molecular and functional identification of a novel photopigment in Pecten ciliary photoreceptors.

The two basic animal photoreceptor types, ciliary and microvillar, use different light-transduction schemes: their photopigments couple to Gt versus Gq proteins, respectively, to either mobilize cyclic nucleotides or trigger a lipid signaling cascade. A third class of photoreceptors has been described in the dual retina of some marine invertebrates; these present a ciliary morphology but operate via radically divergent mechanisms, prompting the suggestion that they comprise a novel lineage of light sensors. In one of these organisms, an uncommon putative opsin was uncovered that was proposed to signal through Go Orthologues subsequently emerged in diverse phyla, including mollusks, echinoderms, and chordates, but the cells in which they express have not been identified, and no studies corroborated their function as visual pigments or their suggested signaling mode. Conversely, in only one invertebrate species, Pecten irradians, have the ciliary photoreceptors been physiologically characterized, but their photopigment has not been identified molecularly. We used the transcriptome of Pecten retina to guide the cloning by polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE) extensions of a new member of this group of putative opsins. In situ hybridization shows selective transcription in the distal retina, and specific antibodies identify a single band of the expected molecular mass in Western blots and distinctly label ciliary photoreceptors in retina sections. RNA interference knockdown resulted in a reduction in the early receptor current-the first manifestation of light transduction-and prevented the prolonged aftercurrent, which requires a large buildup of activated rhodopsin. We also obtained a full-length clone of the α-subunit of a Go from Pecten retina complementary DNA and localized it by in situ hybridization to the distal photoreceptors. Small interfering RNA targeting this Go caused a specific depression of the photocurrent. These results establish this novel putative opsin as a bona fide visual pigment that couples to Go to convey the light signal.

Ciliopathy Protein Tmem107 Plays Multiple Roles in Craniofacial Development

A broad spectrum of human diseases called ciliopathies is caused by defective primary cilia morphology or signal transduction. The primary cilium is a solitary organelle that responds to mechanical and chemical stimuli from extracellular and intracellular environments. Transmembrane protein 107 (TMEM107) is localized in the primary cilium and is enriched at the transition zone where it acts to regulate protein content of the cilium. Mutations in TMEM107 were previously connected with oral-facial-digital syndrome, Meckel-Gruber syndrome, and Joubert syndrome exhibiting a range of ciliopathic defects. Here, we analyze a role of Tmem107 in craniofacial development with special focus on palate formation, using mouse embryos with a complete knockout of Tmem107. Tmem107–/– mice were affected by a broad spectrum of craniofacial defects, including shorter snout, expansion of the facial midline, cleft lip, extensive exencephaly, and microphthalmia or anophthalmia. External abnormalities were accompanied by defects in skeletal structures, including ossification delay in several membranous bones and enlargement of the nasal septum or defects in vomeronasal cartilage. Alteration in palatal shelves growth resulted in clefting of the secondary palate. Palatal defects were caused by increased mesenchymal proliferation leading to early overgrowth of palatal shelves followed by defects in their horizontalization. Moreover, the expression of epithelial stemness marker SOX2 was altered in the palatal shelves of Tmem107–/– animals, and differences in mesenchymal SOX9 expression demonstrated the enhancement of neural crest migration. Detailed analysis of primary cilia revealed region-specific changes in ciliary morphology accompanied by alteration of acetylated tubulin and IFT88 expression. Moreover, Shh and Gli1 expression was increased in Tmem107–/– animals as shown by in situ hybridization. Thus, TMEM107 is essential for proper head development, and defective TMEM107 function leads to ciliary morphology disruptions in a region-specific manner, which may explain the complex mutant phenotype.

Serotonin 5-HT6 receptors affect cognition in a mouse model of Alzheimer\u2019s disease by regulating cilia function

BackgroundSerotonin receptor 5-HT6 is involved in cognition and Alzheimer’s disease (AD) development. However, the mechanism of 5-HT6 in AD pathology is not clear.MethodsSince 5-HT6 is almost exclusively expressed in the primary cilia, using immunostaining we examined the number of cilia in the hippocampus of AD animal model APP/PS1 mice. By overexpressing and knocking down 5-HT6 in the primary cultured hippocampal neurons, we investigated the roles of 5-HT6 in alternating ciliary morphology. Furthermore, 5-HT6 antagonist was applied to confirm its roles in cognition using the Morris water maze test, Y maze, and fear conditioning.ResultsIn the present study, we found that the primary cilia were elongated in the hippocampus of APP/PS1 mice compared with WT mice. 5-HT6 regulated cilia length, influenced cilia and axon initial segment (AIS) morphology, and affected localization of ARL13B and AnkG. We also found that, by changing cilia morphology, the AIS was elongated, branched, and more proximal to the cell body in both WT and APP/PS1 mouse neurons. Alterations of cilia also decreased the axonal length in WT and APP/PS1 neurons. Furthermore, in the water maze test, Y maze, and fear conditioning test, 5-HT6 antagonist SB271046 recovered the cognitive impairment of APP/PS1 mice.ConclusionWe suggest that 5-HT6 plays a critical role in AD development through regulating the morphology and function of neuronal primary cilia, which is possibly related to the AIS and axon alterations in AD development.

Cell Cycle-Related Kinase (CCRK) regulates ciliogenesis and Hedgehog signaling in mice.

The Hedgehog (Hh) signaling pathway plays a key role in cell fate specification, proliferation, and survival during mammalian development. Cells require a small organelle, the primary cilium, to respond properly to Hh signals and the key regulators of Hh signal transduction exhibit dynamic localization to this organelle when the pathway is activated. Here, we investigate the role of Cell Cycle Related kinase (CCRK) in regulation of cilium-dependent Hh signaling in the mouse. Mice mutant for Ccrk exhibit a variety of developmental defects indicative of inappropriate regulation of this pathway. Cell biological, biochemical and genetic analyses indicate that CCRK is required to control the Hedgehog pathway at the level or downstream of Smoothened and upstream of the Gli transcription factors, Gli2 and Gli3. In vitro experiments indicate that Ccrk mutant cells show a greater deficit in response to signaling over long time periods than over short ones. Similar to Chlamydomonas mutants lacking the CCRK homolog, LF2, mouse Ccrk mutant cells show defective regulation of ciliary length and morphology. Ccrk mutant cells exhibit defects in intraflagellar transport (the transport mechanism used to assemble cilia), as well as slowed kinetics of ciliary enrichment of key Hh pathway regulators. Collectively, the data suggest that CCRK positively regulates the kinetics by which ciliary proteins such as Smoothened and Gli2 are imported into the cilium, and that the efficiency of ciliary recruitment allows for potent responses to Hedgehog signaling over long time periods.

Diagnosis of primary ciliary dyskinesia: When and how?

IntroductionPrimary ciliary dyskinesia (PCD) is a rare congenital disorder involving permanent ubiquitous structural and/or functional ciliary abnormalities. MethodsA single-center retrospective study included 56 cases of PCD (respiratory form) out of a cohort of 280 patients with suspected PCD. The main features of history-taking and clinical examination were analyzed, to formulate a pragmatic diagnostic procedure, easy to implement in clinical practice. ResultsChronic respiratory tract infectious symptoms are sensitive but non-specific for the diagnosis of PCD. Nasal brushing for phase-contrast microscopy study of ciliary morphology and activity proved to be a fast, easy, non-invasive, cost-effective and age-independent diagnostic method. In doubtful cases, depending on local availability, further tests are indicated: nasal nitric oxide level, electronic microscopy, genetic study and cell culture. ConclusionsIn suspected PCD, there being no gold standard method of screening and early diagnosis, nasal brushing with ciliary study is contributive, alongside numerous other complementary tests, on condition that the clinician is experienced and results are interpreted in the light of clinical examination and history-taking.

Eyes shut homolog is required for maintaining the ciliary pocket and survival of photoreceptors in zebrafish.

ABSTRACTMutations in the extracellular matrix protein eyes shut homolog (EYS) cause photoreceptor degeneration in patients with retinitis pigmentosa 25 (RP25). Functions of EYS remain poorly understood, due in part to the lack of an EYS gene in mouse. We investigated the localization of vertebrate EYS proteins and engineered loss-of-function alleles in zebrafish. Immunostaining indicated that EYS localized near the connecting cilium/transition zone in photoreceptors. EYS also strongly localized to the cone outer segments and weakly to the rod outer segments and cone terminals in primate retinas. Analysis of mutant EYS zebrafish revealed disruption of the ciliary pocket in cone photoreceptors, indicating that EYS is required for maintaining the integrity of the ciliary pocket lumen. Mutant zebrafish exhibited progressive loss of cone and rod photoreceptors. Our results indicate that EYS protein localization is species-dependent and that EYS is required for maintaining ciliary pocket morphology and survival of photoreceptors in zebrafish.

Mutations in CEP78 Cause Cone-Rod Dystrophy and Hearing Loss Associated with Primary-Cilia Defects

Cone-rod degeneration (CRD) belongs to the disease spectrum of retinal degenerations, a group of hereditary disorders characterized by an extreme clinical and genetic heterogeneity. It mainly differentiates from other retinal dystrophies, and in particular from the more frequent disease retinitis pigmentosa, because cone photoreceptors degenerate at a higher rate than rod photoreceptors, causing severe deficiency of central vision. After exome analysis of a cohort of individuals with CRD, we identified biallelic mutations in the orphan gene CEP78 in three subjects from two families: one from Greece and another from Sweden. The Greek subject, from the island of Crete, was homozygous for the c.499+1G>T (IVS3+1G>T) mutation in intron 3. The Swedish subjects, two siblings, were compound heterozygotes for the nearby mutation c.499+5G>A (IVS3+5G>A) and for the frameshift-causing variant c.633delC (p.Trp212Glyfs(∗)18). In addition to CRD, these three individuals had hearing loss or hearing deficit. Immunostaining highlighted the presence of CEP78 in the inner segments of retinal photoreceptors, predominantly of cones, and at the base of the primary cilium of fibroblasts. Interaction studies also showed that CEP78 binds to FAM161A, another ciliary protein associated with retinal degeneration. Finally, analysis of skin fibroblasts derived from affected individuals revealed abnormal ciliary morphology, as compared to that of control cells. Altogether, our data strongly suggest that mutations in CEP78 cause a previously undescribed clinical entity of a ciliary nature characterized by blindness and deafness but clearly distinct from Usher syndrome, a condition for which visual impairment is due to retinitis pigmentosa.

Evaluation of the Changes in Anterior Segment Morphology with Ultrasound Biomicroscopy after Vitrectomy without Use of Tamponade in Pseudophakic Eyes

Aim: In this study we investigated the changes in anterior segment morphology in pseudophakia patients that underwent ultrasound biomicroscopy (UBM) after pars plana vitrectomy (PPV) operation without use of tamponade. Method: Pseudophakic patients who undergo PPV were enrolled in this prospective study between October 2012 and April 2015. Study included patients in whom intraocular tamponade was not used during PPV operation. UBM measurements were performed both before and 10 days after the operation. Anterior chamber depth (ACD) was measured using axial images of anterior segment. Trabecular meshwork-iris angle (TIA), ciliary body thickness (CBT), sclera thickness (ST), trabecular meshwork-ciliary process distance (T-CPD), iris-ciliary processes distance (I-CPD), and iris thickness (IT) were measured at temporal quadrant based on radial section images of the angle. Values measured before and after the operation were statistically compared with each other. Results: This study included 30 patients (18 females, 12 males) that underwent an operation ultrasound biomicroscopy (UBM) after pars plana vitrectomy (PPV) operation without use of tamponade. Mean age was 69.6 ± 9.1 (55 - 85) years. Eighteen operations occurred on the left eye whereas twelve operations occurred on the right eye. CBT1, CBT2, CBT Max, T-CPD, and I-CPD were significantly decreased after operation when compared with the values of baseline (before) (p = 0.018, p = 0.012, p = 0.001, p = 0.033, p = 0.015, respectively). Other evaluated parameters did not show statistically significant changes after the operation (p > 0.05). Discussion: PPV results in significant changes in ciliary body morphology together with changes in anterior segment parameters in pseudophakic cases.

Comparison of Ciliary Body Anatomy between American Caucasians and Ethnic Chinese Using Ultrasound Biomicroscopy

Purpose: To compare the ciliary body morphology between the Caucasians and Chinese, and to determine the associated anterior segment parameters.Methods: The study groups were comprised of four age- and gender-matched cohorts: American Caucasians, American Chinese, southern and northern mainland Chinese. Ultrasound biomicroscopy (UBM) was performed to assess the following measurements of the ciliary body: trabecular-ciliary process distance (TCPD), trabecular-ciliary process angle (TCA) and ciliary body thickness at 1 mm posterior to the scleral spur (CBT1). A-scan ultrasonography was used to measure lens thickness (LT) and relative lens position (RLP), defined as (ACD + LT/2)/AL.Results: There were 109, 111, 115 and 117 subjects enrolled in the American Caucasian, American Chinese, and southern and northern mainland Chinese groups, respectively. Collectively, the Chinese had smaller TCPD (0.834 ± 0.234 versus 0.940 ± 0.236 mm, p < 0.001) and TCA (75.1 ± 17.9 versus 85.0 ± 21.5, p < 0.001) than Caucasians. No difference was found between the three Chinese subgroups, therefore they were combined as one for the following analyses. After multiple linear regression analysis, adjusting for age, gender, spherical equivalent (SE) and axial length (AL), TCPD and TCA were positively associated with CBT1 and RLP, but negatively correlated with LT in the whole sample. The ethnic Chinese had significantly smaller CBT1 (SRC = −0.177, p < 0.001) and RLP (SRC = −0.254, p < 0.001) than that of Caucasians.Conclusions: Compared with the Caucasians, the Chinese had a thinner ciliary body and a more anteriorly positioned lens, which may contribute to more anteriorly positioned ciliary processes in this population. These findings might help to explain the higher prevalence of angle closure in the Chinese.

Characterization of Primary Cilia Distribution and Morphology During Lactation, Stasis, and Involution in the Bovine Mammary Gland

Primary cilia are small, sensory organelles projecting from virtually all cells and are vital for cellular and tissue function. Their distribution in bovine mammary tissue has not previously been assessed, despite the potential for these organelles to provide specialized perceptive and regulatory functions to this acutely responsive and adaptive gland. The research objectives were to assess ciliary distribution and morphology during active lactation, milk stasis, and early involution using tissue samples obtained following the abrupt cessation of milk removal in nonpregnant, Friesian dairy cows at mid-lactation. Routinely processed tissue sections were obtained at intervals from 6 to 192 hr after the last milking (N = 3 animals per group) and assigned to active lactation (6-12 hr), milk stasis (18-36 hr), and early involution (72-192 hr). Primary cilia were observed in luminal secretory epithelial cells (SECs), myoepithelial cells, and stromal cells following fluorescent immunohistochemistry and confocal microscopy. In SECs, some primary cilia appeared deflected against the apical cell membrane. The proportion of those deflected was greater during milk stasis than active lactation. Data show that primary cilia were suitably placed in three important cell types to potentially coordinate various forms of signal transduction relying on both mechanosensation and chemosensation, according to the physical and physiological state of the gland. Their cell-type distribution and morphology provide new directions in the study of mammary regulation to enhance the understanding of how various mammary-specific cellular responses may be initiated by biochemical or local biophysical factors.

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Cilia are conserved organelles found in many cell types in eukaryotes, and their dysfunction causes defects in environmental sensing and signaling transduction; such defects are termed ciliopathies. Distinct cilia have cell-specific morphologies and exert distinct functions. However, the underlying mechanisms of cell-specific ciliogenesis and regulation are unclear. Here we identified a WD40-repeat (WDR) protein, WDR47/NMTN-1, and show that it is specifically required for ciliogenesis of AWB chemosensory neurons in *C. elegans*. WDR47/NMTN-1 is expressed in the AWB chemosensory neuron pair, and is localized at the basal body (BB) of the AWB cilia. Knockout of *wdr47/nmtn-1* specifically causes abnormal AWB neuron cilia morphology, structural integrity, and induces aberrant AWB-mediated aversive behaviors. We further demonstrate that *wdr47/nmtn-1* deletion results in ciliary ultrastructure defects, and affects movement of intraflagellar transport (IFT) particles and their cargo delivery in AWB neurons. Our results indicate that WDR47/NMTN-1 is essential for AWB neuron ciliary morphology and function, which reveal a novel mechanism for cell-specific ciliogenesis. Since WDR47/NMTN-1 is conserved in mammals, our findings may help understand the process of cell-specific ciliogenesis and provide insights for treating ciliopathies.