The trans-Golgi network golgin, GCC185, is required for endosome-to-Golgi transport and maintenance of Golgi structure

Four mammalian golgins are specifically targeted to the trans-Golgi network (TGN) membranes via their C-terminal GRIP domains. The TGN golgins, p230/golgin-245 and golgin-97, are recruited via the GTPase Arl1, whereas the TGN golgin GCC185 is recruited independently of Arl1. Here we show that GCC185 is localized to a region of the TGN distinct from Arl1 and plays an essential role in maintaining the organization of the Golgi apparatus. Using both small interfering RNA (siRNA) and microRNA (miRNA), we show that depletion of GCC185 in HeLa cells frequently resulted in fragmentation of the Golgi apparatus. Golgi apparatus fragments were dispersed throughout the cytoplasm and contained both cis and trans markers. Trafficking of anterograde and retrograde cargo was analysed over an extended period following GCC185 depletion. Early effects of GCC185 depletion included a perturbation in the distribution of the mannose-6-phosphate receptor and a block in shiga toxin trafficking to the Golgi apparatus, which occurred in parallel with the fragmentation of the Golgi ribbon. Internalized shiga toxin accumulated in Rab11-positive endosomes, indicating GCC185 is essential for transport between the recycling endosome and the TGN. In contrast, the plasma membrane-TGN recycling protein TGN38 was efficiently transported into GCC185-depleted Golgi apparatus fragments throughout a 96-h period, and anterograde transport of E-cadherin was functional until a late stage of GCC185 depletion. This study demonstrated (i) a more effective long-term depletion of GCC185 using miRNA than siRNA and (ii) a dual role for the GCC185 golgin in the regulation of endosome-to-TGN membrane transport and in the organization of the Golgi apparatus.     

Developmental expression of dermatan sulfate proteoglycans in the elastic bovine nuchal ligament.

The nuchal ligament of bovines is a useful system in which to study elastic fibre formation since it contains up to 83% elastin and undergoes a period of rapid elastinogenesis during the last trimester of fetal development and in the first four post-natal months. To identify proteoglycans (PGs) which may be involved in this process we initially investigated changes in the glycosaminoglycan (GAG) profiles during nuchal ligament development. In contrast to the collagenous Achilles tendon, nuchal ligament exhibited: (a) elevated hyaluronan (HA) levels in the peak period of elastin-associated microfibril (fibrillin) synthesis (130-200 days) which precedes elastinogenesis; and (b) markedly increased synthesis of a glucuronate-rich copolymeric form of dermatan sulfate (DS) in the period corresponding to elastin formation (200-270 days). Analysis of DSPGs isolated from 230-day nuchal ligament showed that this copolymer was predominantly associated with a glycoform of biglycan which was specifically elevated at this stage in development. This finding was consistent with Northern blot analysis which showed that steady-state biglycan mRNA levels increased significantly during the elastinogenic period. In contrast, the mRNA levels for decorin, the only other DSPG detected in this tissue, declined rapidly after 140 days of fetal development. In conclusion, the results suggest that HA may play a role in microfibril assembly and that a specific glycoform of biglycan may be associated with the elastinogenic phase of elastic fibre formation.     

Unsuspected hypercalcemia among adults in hospital.

Physicians usually fail to recognize an abnormal serum calcium level and often do not arrive at a definite conclusion about its cause. Yet hypercalcemia, even when minimal, indicates potentially serious underlying disease. This review of the records of about 12 000 adults in hospital showed a prevalence of unsuspected hypercalcemia of 1.4%, a 24% rate of recognition of the abnormal serum level and a 71% rate of correct diagnosis (though often presumptive) of the underlying cause by the attending physician, and a possible prevalence of unsuspected hyperparathyroidism of 0.3%. Because of other considerations, however, the prevalence estimates are probably for too low.     

Anther opening, pollen biology and stigma receptivity in the long blooming species, Parietaria judaica L. (Urticaceae)

Background and aimsThis paper is about pollen ecophysiology, anther opening, pollen dispersal and the timing of the male and female phases in Parietaria judaica (Urticaceae).MethodsEcophysiological (effects of different relative humidities (RHs) and osmotic relationships) and cytological methods (stigmatic receptivity, pollen viability, histology and histochemistry) were used to determine pollen and pollination features during the long blooming period of this species.Key resultsPollen is dispersed by rapid uncurling of the filament and anther opening. The filament and anther lack cells with lignified wall thickenings, which are usually responsible for anther opening and ballistic pollen dispersal. Instead, dispersal is the result of the sudden movement of the filament. Pollen is of the partially hydrated type, i.e. with a water content greater than 30% at shedding, and readily loses water, and hence viability, at low RH. Pollen carbohydrate reserves differ with season. Starchless grains germinate quickly and are less subject to water loss. Flowers are protogynous, pollen release occurring only after complete cessation of the female phase within an inflorescence.ConclusionParietaria has partially hydrated pollen which differs from typical pollen of this type because of its reduced size and the absence of callose. Because of its low water content at the time of shedding it survives better at higher RH. Dispersal and pollination are adapted to pollen features.     

Road Casualties among Birds

The Stone Curlew, restricted in its distribution to relatively small areas in south and east England, is a declining species in many localities. Data of the B.T.O. Nest Records and Ringing Schemes have been used to study some of the pressures affecting this species both during and outside the breeding season.